SCF Receptor

SCF Receptor Diana Linnekin1,* and Jonathan R. Keller 2 1

Basic Research Laboratory, Division of Basic Sciences, National Cancer Institute±Frederick Cancer Research & Development Center, Frederick, MD 21702, USA 2 Intramural Research and Support Program, SAIC Frederick, National Cancer Institute±Frederick Cancer Research & Development Center, Frederick, MD 21702, USA * corresponding author tel: 301-846-5631 fax: 301-846-6641, e-mail: [email protected] DOI: 10.1006/rwcy.2000.20003.

SUMMARY The stem cell factor receptor (SCFR) is a member of the receptor tyrosine kinase superfamily. It is expressed on a variety of cell types, including hematopoietic cells, germ cells, neural cells, melanocytes, and the interstitial cells of Cajal in the intestine. The absence of functional murine SCFR is lethal in utero. Mutations resulting in reduced expression, or function, of the SCFR lead to abnormalities in hematopoiesis, pigmentation, and reproduction in mice. Inappropriate expression or activation of the SCFR is associated with a variety of diseases, including mastocytosis in humans. Further, downregulation of the SCFR may play a role in metastatic melanoma. The importance of the SCFR in both normal and pathophysiology has led to extensive study of its mechanism of action. Recent work indicates that the SCFR activates multiple signal transduction pathways, including the Ras/ Raf/MAP kinase cascade, the JAK/STAT pathway, phosphatidylinositol 3-kinase, and Src family members. This review will provide an overview of the structure, biology, and mechanism of action of the SCFR.

BACKGROUND

Discovery As early as 1929, naturally occurring mutations in mice resulting in profound impairment in pigmentation, reproduction, and blood cell development were observed by geneticists. Subsequent work found this phenotype resulted from alterations in either the white spotting (W) or steel (Sl) locus. In 1979,

Elizabeth Russell proposed that the gene product of the white spotting locus was a receptor and that the gene product of the steel locus was its cognate ligand (Russell, 1979). In 1986, v-kit, an oncogene isolated from Hardy±Zuckerman feline sarcoma virus, was cloned and characterized (Besmer et al., 1986). Within 2 years both the human and murine forms of the proto-oncogene c-Kit had been cloned (Yarden et al., 1987; Qiu et al., 1988). c-Kit was subsequently mapped to the white spotting locus and in 1990 the gene product of the steel locus was cloned and found to be the ligand for c-Kit (Chabot et al., 1988; Geissler et al., 1988; Huang et al., 1990; Zsebo et al., 1990). Names for the ligand include steel factor, stem cell factor, c-Kit ligand, and mast cell growth factor.

Alternative names The stem cell factor receptor (SCFR) is also known as c-Kit and has been designated CD117.

Structure The SCFR is a receptor tyrosine kinase (RTK) and is related to the receptors for platelet-derived growth factor (PDGF) and colony-stimulating factor 1 (CSF-1). The full-length murine protein consists of 975 amino acids and the human protein is 976 amino acids (Yarden et al., 1987; Qiu et al., 1988). The extracellular domain consists of approximately 500 amino acids and is followed by a series of hydrophobic amino acids that span the cell membrane. The intracellular region is slightly less than 400 amino acids long and consists of a 30 amino acid

1914 Diana Linnekin and Jonathan R. Keller juxtamembrane region, a catalytic domain divided into two regions by an insert of 77 amino acids and a 50 amino acid carboxyl tail (Figure 1). An alternately spliced form of the SCFR, termed c-KitA in the literature, contains a four-codon insert in the extracellular domain (Reith et al., 1991). In addition, a 24 kDa truncated form of the SCFR consisting of a portion of the second catalytic domain and the carboxyl tail is expressed in spermatids (Rossi et al., 1992).

Main activities and pathophysiological roles Early studies of white spotting and steel mice, as well as subsequent work since the cloning of SCF and the SCFR, have shown that SCF is critical for the survival and development of stem cells involved in hematopoiesis, pigmentation, and reproduction (Galli et al., 1994; Broudy, 1997; Lyman and Jacobsen, 1998; Ashman, 1999). More recently, the SCFR has been found to be important in development of the interstitial cells of Cajal in the intestine, and in certain learning functions in the hippocampal region of the brain (Huizinga et al., 1995; Motro et al., 1996). A comprehensive overview of SCF and its receptor in relation to all of the physiological target tissues was written in 1994 by Galli and coworkers. The following section will summarize the early literature and highlight recent findings relating to the activities and pathophysiological roles of the SCFR in different organ systems. Reviews of the role of the SCFR in hematopoiesis have recently been published (Galli et al., 1994; Broudy, 1997; Lyman and Jacobsen, 1998; Ashman, 1999). The role of the SCFR in the ontogeny of murine hematopoietic progenitor cells is illustrated by the dramatic anemias found in steel and white spotting mice. These animals are characterized by macrocytic anemia, decreases in myeloid and erythroid progenitors, and mast cell deficiencies (Russell, 1979). The ablation of CFU-S, CFU-GM, CFU-IL-3 and CFU-M in adult mice injected with ACK-2, an antibody that blocks the murine SCFR, demonstrates the important role of the SCFR in erythro- and myelopoiesis in adult mice as well (Ogawa et al., 1991). Expression and function of the SCFR have been examined extensively in relation to hematopoietic disease in humans. One disease in which the SCFR likely plays a role is mastocytosis. Reviews dealing with the SCFR and mastocytosis have recently been published (Tsujimura, 1996; FoÈdinger and Mannhaller, 1997; Pignon, 1997; Vliagoftis et al., 1997). In brief,

Metcalfe and coworkers found that patients with mastocytosis with associated hematological disorder have a mutation in the SCFR (substitution of aspartic acid 816 with valine, D816V) that results in constitutive activation (Nagata et al., 1995). A patient with aggressive systemic mastocytosis with the identical mutation in mast cells was also reported by Longley et al. (1996). In all patients described to date, the mutation in the SCFR is somatic. Interestingly, mutation of the analogous amino acid in the RTKs Ret and Met results in constitutive activation and is associated with multiple endocrine neoplasia type 2B and papillary renal carcinomas, respectively (Hofstra et al., 1994; Schmidt et al., 1997). Since these initial reports, a number of groups have reported the D816V mutation in patients with different forms of mastocytosis (Afonja et al., 1998; BuÈttner et al., 1998; Worobec et al., 1998; Longley et al., 1999). Recently, other gain-of-function mutations in the SCFR have been identified in patients with mastocytosis. These include mutations at codons 820 and 560 (Pignon et al., 1997; BuÈttner et al., 1998). In dogs with mastocytosis, mutations in the juxtamembrane region of the SCFR have also been found (Ma et al., 1999). A summary of c-Kit mutations found in patients with mastocytosis is given in Table 1. The relationship of the SCFR to leukemia and lymphoma in humans has also been studied rigorously. These data have recently been reviewed (Galli et al., 1994; Hassan and Zander, 1996; Broudy, 1997; Sperling et al., 1997; Escribano et al., 1998; Lyman and Jacobsen, 1998). In summary, the SCFR is expressed on blast cells from most patients with acute myeloid leukemia (AML), although it is unknown if this plays a role in the onset and progression of the disease. The frequency of SCFR expression on cells from patients with chronic myelogenous leukemia (CML) is lower; however, studies in cell lines suggest that p210bcr/abl binds and activates the SCFR (Hallek et al., 1996). In addition, several proteins phosphorylated in response to SCF are constitutively phosphorylated in CML cells (Wisniewski et al., 1996). These include Dok, a 62 kDa protein, and the adapter protein CRKL (Carpino et al., 1997; Sattler et al., 1997; Yamanashi and Baltimore, 1997; Sattler and Salgia, 1998). In acute lymphoid leukemia (ALL), the SCFR is expressed on malignant lymphoid cells from T-ALL patients while SCFR expression on cells from patients with B-ALL is quite rare. The SCFR may also be a marker for certain types of biphenotypic leukemias. The importance of SCFR in development of melanocytes is highlighted by the abnormal pigmentation found when expression is reduced, or the kinase activity is impaired. In both mice and pigs,

SCF Receptor 1915 Table 1 Activating mutations in the SCFR associated with diseases Region

Amino acid residue(s)

Diseasea

Species

Reference

Extracellular domain

D52N

Myeloproliferative disorder

Human

Kimura et al., 1997

Juxtamembrane

Deletion 559, 560

GIST

Human

Hirota et al., 1998

Juxtamembrane

Deletion 551±555

GIST

Human

Juxtamembrane

V559D

GIST

Human

Juxtamembrane

K550I, Deletion 551±555

GIST

Human

Juxtamembrane

Deletion 550±558

GIST

Human

Juxtamembrane

Deletion 551±554, V555L

GIST

Human

Juxtamembrane

Deletion 556±560

GIST

Human

Juxtamembrane

Q556H, Deletion 557±572

GIST

Human

Juxtamembrane

Deletion 560

GIST

Human

Juxtamembrane

Deletion 557±575

GIST

Human

Juxtamembrane

Deletion 570±576

GIST

Human

Juxtamembrane

V560D

GIST

Human

Juxtamembrane

12 codon insert between 574 and 575

GIST

Human

Juxtamembrane

Deletion 556±557

Mastocytoma

Canine

Moskaluk et al., 1999

Ma et al., 1999

Juxtamembrane

Deletion 558

Mastocytoma

Canine

Juxtamembrane

W556R

Mastocytoma

Canine

Juxtamembrane

L575P

Mastocytoma

Canine

Juxtamembrane

V560G

Mastocytosis

Human

BuÈttner et al., 1998

Catalytic domain

D816V, D816Y, D816F

Mastocytosis

Human

Nagata et al.,1995; Longley et al., 1996; Afonja et al., 1998; Worobec et al., 1998; BuÈttner et al., 1998; Longley et al., 1999

Catalytic domain

D820G

Aggressive mast cell disease

Human

Pignon et al., 1997

a

GIST designates gastrointestinal stromal cell tumor.

abnormalities in the SCFR are associated with a characteristic pattern of white spots occurring predominantly on the ventral trunk, head, tail, and feet (Russell, 1979; Marklund et al., 1998). Similarly, in humans, autosomal dominant piebaldism results from mutations in the SCFR (Fleischman et al., 1991; Giebel and Spritz, 1991; Spritz, 1994, 1997; Boissy and Nordlund, 1997). Expression of functional SCFR appears to be critical at several points during the development and maturation of melanocytes (Nishikawa et al., 1991; Okura et al., 1995; WehrleHaller and Weston, 1995). This includes proliferation of melanoblasts in the mesoderm leading to entry into the epidermal layers during embryogenesis. In

addition, the SCFR plays a role in the replenishment of melanocytes involved in hair pigmentation in adults. The important role of SCFR in melanocyte development has led investigators to examine involvement in melanoma. While SCFR has been reported in some melanoma tissue, as well as in melanoma-derived cell lines, many investigators have found that SCFR is either absent, or expressed at reduced levels, as compared with normal melanocytes (Funasaka et al., 1992; Lassam and Bickford, 1992; Gutman et al., 1994; Mattei et al., 1994; Ohashi et al., 1996; Montone et al., 1997). Huang and coworkers found that infection of a highly metastatic melanoma cell line with a retroviral expression vector containing

1916 Diana Linnekin and Jonathan R. Keller the SCFR reduced metastatic potential (Huang et al., 1996). They also found that SCF induced apoptosis of these cells. Thus, downregulation of the SCFR in melanoma may promote expansion of transformed cells due to escape from SCF-induced apoptosis. Recently, a strong correlation between loss of expression of the AP-2 transcription factor and downregulation of the SCFR in melanoma-derived tissues has been demonstrated (Huang et al., 1998). The data examining the SCFR in relation to metastatic melanoma have recently been reviewed (Lu and Kerbel, 1994; Luca and Bar-Eli, 1998). The receptor for SCF can be detected in primordial germ cells (PGCs) as early as day 7 of embryonic development. SCF acts as a viability factor for PGCs and also induces limited increases in proliferation (Donovan, 1994). Similar to hematopoietic cells, SCF in combination with other growth factors elicits synergistic increases in proliferation of PGCs. In developing sperm, the SCFR has been detected in type A spermatogonia as well as primary spermatocytes (Morrison-Graham and Yoshiko, 1993; Loveland and Schlatt, 1997). A truncated form of the SCFR, consisting of the second portion of the catalytic domain, as well as the C-terminal tail, is expressed in spermatids (Rossi et al., 1992). Interestingly, injection of this form of SCFR into mouse oocytes results in parthenogenesis (Sette et al., 1997). The SCFR has also been detected in Leydig cells in the testicular interstitium. The role of SCFR in the development of sperm in adults was recently reviewed by Loveland and Schlatt (1997). The importance of SCFR in male fertility is illustrated by male sterility in both white spotting and steel mice. In addition, SCFR may also be a marker for certain forms of testicular cancer (Rukstalis, 1996; Donovan et al., 1998). In an experimental model, nearly 100% of transgenic mice expressing papillomavirus type 16 (HPV16) develop testicular cancer (Kondoh et al., 1995; Li et al., 1996). Introduction of either the steel dickie mutation encoding soluble SCF, or white spotting mutants, with impaired SCFR kinase activity, dramatically reduces tumor volume in these animals (Kondoh et al., 1995; Li et al., 1996). In postembryonic females, SCFR is expressed on both oocytes and theca cells, while the SCF ligand is expressed on granulosa cells (Driancourt and Thuel, 1998). SCF interaction with SCFR plays a role in early follicular development, formation of preantrum follicules and ovarian follicular maturation (Yoshida et al., 1997). In contrast, formation and survival of primordial follicles, antral follicular development, ovulation, and follicular luteinization do not require the SCFR. In addition, activation of the SCFR on oocytes by SCF on granulosa cells may play a role in meiotic arrest of oocytes. Further, downregulation of

SCF by gonadotropins may relieve meiotic arrest (Laitinen et al., 1995; Ismail et al., 1996, 1997). Both SCF and the SCFR are presently being examined for potential involvement in premature ovarian failure in humans (Bondy et al., 1998). In the gastrointestinal tract, the SCFR is expressed on the interstitial cells of Cajal (ICC) (Torihashi et al., 1995). These cells play a critical role in the pacemaker activity of the gut. Interestingly, in white spotting viable mice, ICC cells are absent and pacemaker activity is dramatically reduced (Huizinga et al., 1995). In addition, these animals may be more susceptible to necrotizing enterocolitis (Yamataka et al., 1998). Humans with piebaldism resulting from mutations in the SCFR have also been reported to have defects in intestinal function (Giebel and Spritz, 1991). Many gastrointestinal stromal tumors (GISTs) express a constitutively active form of the SCFR resulting from mutations in the juxtamembrane region (Table 1). The association of mutations in c-Kit with GISTs has recently been reviewed (Kitamura et al., 1998; Chan, 1999). As described above, another constitutively active SCFR mutant, D816V, is found in patients with mastocytosis and associated hematological disorders. Interestingly, some of these patients also have gastrointestinal problems. Another organ system where the SCFR is expressed extensively is the central nervous system (CNS). Detailed descriptions of SCFR localization in the adult CNS have previously been published and reviewed (Hirota et al., 1992; Morii et al., 1992; Hamel and Westphal, 1997; Zhang and Fedoroff, 1997). Heterozygote steel dickie mice have normal long-term potentiation but impaired configural learning (Motro et al., 1996). This demonstrates an important role for signaling of membrane-bound SCF in some aspects of brain function. With regard to cancers of the CNS, the SCFR has been observed in some, but not all, neuroblastoma and glioblastoma cell lines (Hamel and Westphal, 1997). SCFR is expressed on Nf1-deficient Schwann cell lines and may play a role in Schwann cell hyperplasia observed in neurofibromin-deficient patients (Badache et al., 1998). Transgenic mice expressing portions of the SCFR promoter fused to the large T antigen of SV40 develop neuroendocrine tumors at high frequencies (Bosse et al., 1997). These findings suggest CNSspecific regulatory sequences in the 50 region of the SCFR gene. The SCFR may also be involved in nonmalignant diseases of the CNS. A recent study by He and coworkers (1997) found that infection of cultured fetal brain cells with HIV-1 induced expression of the SCFR. This was associated with high levels of apoptosis in astrocytes, suggesting that inappropriate expression of the SCFR in the CNS

SCF Receptor 1917 could be involved in neurological disorders associated with AIDS. The Kit gene product has also been associated with other forms of cancer. The v-Kit oncogene is a component of Hardy-Zuckerman strain of feline sarcoma virus (Besmer et al., 1986). Approximately 70% of all small cell lung carcinomas (SCLC) coexpress the SCFR and SCF inappropriately (Hibi et al., 1991; Sekido et al., 1991; Plummer et al., 1993; Rygaard et al., 1993). Thus, an autocrine loop may play a role in the etiology of this disease (Krystal et al., 1996). Similarly, in one study, the SCFR and SCF were coexpressed in 9 of 11 breast tumors, and in 7 of 13 cell lines derived from breast tumors (Hines et al., 1995). Downregulation of the SCFR in the epithelial cells of both breast cancer and thyroid cancer tissue has also been reported (Natali et al., 1995; Chui et al., 1996). Thus, similar to its actions on melanocytes, SCF may play a role in promoting apoptosis in these tissues and downregulation of SCFR may promote transformation. In cell lines derived from colon carcinoma tissue, truncated forms of the SCFR encoding a portion of the second catalytic domain have been identified (Toyota et al., 1994; Takaoka et al., 1997). What, if any, role of these forms of the SCFR play in colon carcinoma is unknown. The SCFR has also been found in cell lines derived from a variety of other types of cancers (Natali et al., 1992; Turner et al., 1992; Matsuda et al., 1993; Inoue et al., 1994; Ridings et al., 1995; DiPaola et al., 1997).

GENE

Accession numbers The cDNA sequence for the human SCFR (GenBank X06182) was reported in 1987 and for the murine SCFR (GenBank Y00864) in 1988 (Yarden et al., 1987; Qiu et al., 1988). The genomic sequence for both murine and human SCFR have also been reported (Andre et al., 1992; Giebel et al., 1992; Gokkel et al., 1992; Vandenbark et al., 1992; Yasuda et al., 1993). The GenBank accession number for the complete coding sequence of the human SCFR is U63834 and for portions of the murine gene, including the 50 regulatory region, is X65998. The human SCFR gene is located on chromosome 4 and the murine gene is on chromosome 5 (Yarden et al., 1987). The c-Kit gene is over 70 kb in size and has 21 exons (Andre et al., 1992; Giebel et al., 1992; Gokkel et al., 1992; Vandenbark et al., 1992; Yasuda et al., 1993). The organization is highly conserved between the mouse and human genome. The c-Kit transcript is 5.2 kb and alternately spliced forms have been reported.

PROTEIN

Accession numbers GenBank: Human SCFR: 1817733 Murine SCFR: 125473

Description of protein The SCFR belongs to the type III family of receptor tyrosine kinases (Yarden and Ullrich, 1988). A model depicting the structural domains of the SCFR is shown in Figure 1. The extracellular domain is organized into a series of five Ig-like regions. The first three Ig-like regions bind SCF while the fourth may play a role in either forming or stabilizing SCFR homodimers (Lev et al., 1992a; Blechman et al., 1993a, 1995; Lev et al., 1993; Blechman et al., 1993b). The role of the fifth Ig-like region is unknown. The cytoplasmic domain of the SCFR can be divided into five regions. The 30 amino acids distal to the

Figure 1 Structural domains of the SCFR. The transmembrane domain divides the SCFR into an intracellular and extracellular region. The intracellular region consists of five immunoglobulin-like domains. The first three bind SCF, the fourth domain may play a role in forming or stabilizing receptor dimers, and the function of the fifth is unknown. The 30 amino acids distal to the transmembrane domain form the juxtamembrane region. The catalytic domain is divided into two by a kinase insert and the last 50 amino acids comprise the kinase tail.

Immunoglobulin-like domain 1–3: binds ligand Extracellular Immunoglobulin-like domain 4: potential role in receptor dimerization Immunoglobulin-like domain 5: unknown function Transmembrane region Juxtamembrane region Catalytic domain 1 Kinase insert Intracellular Catalytic domain 2 Tail

1918 Diana Linnekin and Jonathan R. Keller transmembrane region comprises the juxtamembrane region. The catalytic domain is divided by a 77 amino acid insert (Figure 1). The first catalytic domain contains the ATP-binding region while the second catalytic domain has a number of possible autophosphorylation sites. Lastly, there is a carboxyl-tail of approximately 50 amino acids.

Relevant homologies and species differences The sequences of the human and murine SCFR were published in 1987 and 1988 respectively (Yarden et al., 1987; Qiu et al., 1988). The amino acid homology is 81.9%. Although the human and murine forms of the SCFR are relatively similar, there are significant differences in ligand specificity. Both murine and human SCFR bind and are activated by murine SCF. In contrast, human SCF binds the murine receptor with extremely low affinity and does not have significant biological activity on murine cells. Other species for which the SCFR sequence is known include rat, bird, goat, pig, cow, cat, and dog (Tsujimura et al., 1991; Sasaki et al., 1993; Kubota et al., 1994; Zhang and Anthony, 1994; Herbst et al., 1995a; Tanaka et al., 1997; Ma et al., 1999).

Affinity for ligand(s) SCF binds the SCFR with an affinity of 50±200 pM in both hematopoietic and nonhematopoietic target tissue (Abkowitz et al., 1992; Turner et al., 1992).

Cell types and tissues expressing the receptor The embryonic expression patterns of the SCFR have previously been published (Matsui et al., 1990; Orr-Urtreger et al., 1990; Keshet et al., 1991; Motro et al., 1991). The following section will summarize expression of the SCFR in adult tissues. SCFR expression in hematopoietic cells was recently reviewed by Lyman and Jacobsen (1998). In brief, the SCFR is expressed on stem cells, pluripotential progenitor cells and more differentiated progenitor cells including those leading to the myeloid, lymphoid, erythroid and megakaryocytic lineages. SCFR expression levels vary with the differentiation of murine marrow cells. Early progenitors can be divided into SCFRdull or SCFRbright populations and in general,

SCFR expression declines as progenitor cells differentiate. Certain terminally differentiated lineages do express the SCFR. These include mast cells, activated platelets and a subset of NK cells. SCFR is also expressed in numerous cell lines isolated from hematopoietic cells. Table 2 is a summary of some of these lines, the species of origin, the hematopoietic lineage, if known, and whether the cells require hematopoietic growth factors for propagation in culture. In the postnatal female, SCFR is expressed in both oocytes and theca cells (Draincourt and Truel, 1998). In the postnatal male, full-length SCFR is expressed in type A spermatogonia as well as primary spermatocytes (Morrison-Graham and Yoshiko, 1993; Loveland and Schlatt, 1997). In addition, a truncated form has been found in spermatids. The SCFR is also expressed in the testicular interstitium in the Leydig cells. SCFR is expressed throughout the CNS. Descriptions of its localization have previously been published (Hirota et al., 1992; Morii et al., 1992; Hamel and Westphal, 1997; Zhang and Fedoroff, 1997). Other tissues that express the SCFR in adults are the interstitial cells of Cajal in the intestine, the oval and bile epithelial cells in the liver, alveolar, ductal, and epithelial cells in the breast and the glandular epithelial cells in the parotid, dermal sweat, and salivary glands as well as the esophagus (Lammie et al., 1994; Torihashi et al., 1995; Fujio et al., 1996). SCFR has also been found in human endometrium and placenta during pregnancy (Saito et al., 1994; Kauma et al., 1996).

Regulation of receptor expression Surface expression of the SCFR is regulated through multiple mechanisms. Similar to other receptors, ligand-induced activation rapidly induces internalization (Yee et al., 1993, 1994a; Miyazawa et al., 1994; Shimizu et al., 1996; Gommerman et al., 1997; Broudy et al., 1998). While both soluble and membranebound SCF induce internalization of the SCFR, the kinetics are more rapid in response to the soluble form (Miyazawa et al., 1995). One mechanism mediating internalization involves interaction of the SCFR with clathrin, incorporation into clathrin-coated pits and subsequent degradation, in part, by lysosomal proteases (Gommerman et al., 1997). A second mechanism mediating downregulation of activated SCFR is through rapid ubiquination and degradation though the proteasome proteolytic pathway (Miyazawa et al., 1994).

SCF Receptor 1919 Table 2

Hematopoietic cell lines that express the SCFR

Designation

Species

Lineage

Growth factordependence

HEL

Human

Erythroid

No

LAMA

Human

Erythroid

No

K562YO

Human

Erythroid

No

OCIM1

Human

Erythroid

No

OCIM2

Human

Erythroid

No

JK-1

Human

Erythroid

No

HMC1

Human

Mast cell

No

CMK

Human

Megakaryoblastic

No

DAMI

Human

Megakaryoblastic

No

MEG-01

Human

Megakaryoblastic

No

Mo7e

Human

Megakaryoblastic

Yes

HML-2

Human

Megakaryoblastic

Yes

MB-02

Human

Erythroid

Yes

a

TF-1

Human

Erythroid

Yes

UT-7

Human

Erythroid

Yes

F-36P

Human

Erythroid

Yes

Human

Myeloid

Yes

AML193

Human

Myeloid

Yes

P815

Mouse

Mastocytoma

No

FMA3

Mouse

Mastocytoma

No

SKT6

Mouse

Erythroid

No

B6M

Mouse

Mast cell

Yes

MC-9

Mouse

Mast cell

Yes

R6-X

Mouse

Mast cell/meg.

Yes

FDC-P1

Mouse

Progenitor

Yes

FDCP-Mix A4

Mouse

Progenitor

Yes

B6SUtA1

Mouse

Myeloid

Yes

NSF-60

Mouse

Myeloid

Yes

HCD-57

Mouse

Erythroid

Yes

GF-D8 a

a

Clones of these cell lines have been reported that do not express the SCFR.

Expression of the SCFR can also be regulated by other cytokines and growth factors (Galli et al., 1994; Broudy et al., 1998; Lyman and Jacobsen, 1998). In brief, treatment of hematopoietic progenitor cells, as well as cell lines, with TGF , TNF, or IL-4 reduces expression of the SCFR (Sillaber et al., 1991; Khoury et al., 1994; Nilsson et al., 1994; Rusten et al., 1994; Heinrich et al., 1995; Jacobsen et al., 1995). One mechanism mediating TGF -induced downregulation of the SCFR is reduction in mRNA stability

(de Vos et al., 1993; Dubois et al., 1994; Heinrich et al., 1995). IFN also downregulates expression of SCFR on erythroid progenitor cells (Taniguchi et al., 1997). There are varied reports on the effects of IL-3 on SCFR expression. One group found that IL-3 and GM-CSF had no effect on expression of SCFR mRNA in the megakaryoblastic cell line Mo7e (Hu et al., 1994). Treatment of mast cells, a mast cell line, or a myeloid cell line with IL-3 downregulated SCFR mRNA as well as protein levels (Welham and

1920 Diana Linnekin and Jonathan R. Keller

Release of soluble receptors Another mechanism for regulation of cell surface expression of SCFR is the release of soluble SCFR from the cell surface through proteolytic cleavage and release of the extracellular domain. Soluble SCFR is generated by umbilical endothelial cells, hematopoietic cell lines, and mast cells (Brizzi et al., 1994a; Turner et al., 1995). Further, soluble SCFR has been detected at relatively high concentrations in human plasma (Wypych et al., 1995). Treatment of cells with either phorbol myristate acetate or calcium ionophore results in release of soluble SCFR into the supernatant (Asano et al., 1993; Yee et al., 1993). While the biological role of soluble SCFR is unknown, it is capable of binding SCF and antagonizing SCFmediated responses.

SIGNAL TRANSDUCTION

Associated or intrinsic kinases

Figure 2 SCF activates multiple signaling pathways. Ligand-induced increases in tyrosine phosphorylation of the SCFR recruit multiple signaling components to the receptor complex. These include Grb2, Shc, and SHP2, proteins involved in regulation of the Ras/Raf/ MAP kinase cascade. The 85 kDa regulatory subunit of PI-3 kinase also binds phosphorylated SCFR and results in activation of the catalytic subunit of PI-3 kinase. Src family members, JAK2, Tec, PLC and STAT1 also associate with the SCFR and are activated by SCF.

F

Shp2 ily am rc f

b2

Gr

Shc

s

So

Ras

c-Raf-1

S

l Cb

Cytoplasmic signaling cascades Similar to other receptor tyrosine kinases, SCFR activates multiple signal transduction components that couple to a variety of biochemical pathways. Among these are phosphatidylinositol 3-kinase (PI-3 kinase), Src family members, the JAK/STAT pathway and the

SC

The SCFR is a receptor tyrosine kinase. Binding of SCF induces rapid increases in homodimerization of the receptor followed by increases in the SCFR autophosphorylation activity in multiple lineages of cells including hematopoietic, melanocytes, and spermatozoa (Blume-Jensen et al., 1991; Funasaka et al., 1992; Lev et al., 1992a, 1992b; Philo et al., 1996; Lemmon et al., 1997; Feng et al., 1998). The critical role of the SCFR kinase activity in responses mediated by SCF is illustrated by the severe phenotype of white spotting mice expressing SCFR lacking kinase activity. In addition, gain-of-function mutations in the SCFR lead to oncogenic forms of the receptor and have also been found in patients with gastrointestinal stromal tumors (GISTs) and mastocytosis (Nagata et al., 1995; Longley et al., 1996; Hirota et al., 1998; Moskaluk et al., 1999).

Ras/Raf/MAP kinase cascade. A model of signaling components and pathways activated by SCF is shown in Figure 2. A review of signaling pathways activated by SCF in hematopoietic cells has recently been published (Linnekin, 1999). The following summarizes each of these pathways in relation to SCF signaling. PI-3 kinase is a heterodimer composed of an 85 kDa regulatory subunit and a 110 kDa catalytic subunit involved in the signaling pathways of a variety of growth factors and cytokines (Shepherd et al., 1998). Studies in both transfected fibroblasts, as well as hematopoietic cells, have shown that SCF induces increases in SCFR-associated PI-3 kinase activity. SCF also induces increases in tyrosine phosphorylation of the 85 kDa subunit of PI-3 kinase (Lev et al., 1991; Reith et al., 1991; Rottapel et al., 1991). SCFR mutants lacking kinase activity do not activate PI-3 kinase in response to SCF, while cells expressing a constitutively active SCFR mutant have increased levels of receptor-associated PI-3 kinase activity (Rottapel et al., 1991). Thus, activation of PI3 kinase by the SCFR correlates directly with SCFR catalytic activity. Tyrosine 719 of murine SCFR (Y721 of human SCFR) interacts with the SH2 domain of PI-3 kinase.

SCF

Schrader, 1991). Similar results were found after treatment of a mast cell line with EPO or GM-CSF, but not IL-4.

c-

Cr

k1

p85 P13K JAK2 Stat 1

AKT

p110 p13K

MAP kinase cascade Tec

PLC

SCF Receptor 1921 Studies of SCFR mutants containing phenyalanine at residue 719 (Y719F) suggest that PI-3 kinase has multiple roles in SCF-mediated responses. These include adhesion to fibronectin, as well as membrane ruffling and actin assembly in mast cells (Serve et al., 1995; Vosseller et al., 1997). Partial inhibition of SCFmediated proliferation and survival have also been observed in murine mast cells expressing Y719F SCFR (Serve et al., 1995; Vosseller et al., 1997). In DA-1 cells, a murine myeloid cell line, PI-3 kinase plays a role in trafficking of the SCFR through endocytic pathways (Gommerman et al., 1997). In the myeloid progenitor cell line FDC-P1, PI-3 kinase may also play a role in differentiation mediated by the SCFR (Kubota et al., 1998). Lastly, SCF-induced activation of the serine threonine kinase Akt is impaired in fibroblast cell lines expressing Y721F human SCFR. This correlates with a partial impairment in capacity of SCF to prevent apoptosis (BlumeJensen et al., 1998). Thus, the role of PI-3 kinase in SCF-mediated responses depends on cell lineage. SCF also activates the JAK/STAT pathway (Linnekin et al., 1997a). Studies from two groups have shown that JAK2 associates with the SCFR and that SCF activates JAK2 (Brizzi et al., 1994b; Weiler et al., 1996). Studies with antisense oligonucleotides suggest that JAK2 is required for maximal proliferation in response to SCF. SCF also induces association of STAT1 with the SCFR, increases in STAT1 tyrosine phosphorylation and increases in STAT1 DNA-binding activity in multiple cell lines (DeBerry et al., 1997). In addition to STAT1, SCF also activates STAT5, and induces serine phosphorylation of STAT3 (Gotoh et al., 1996; Ryan et al., 1997). In contrast to these findings, several investigators have reported that SCF has no effect on JAK or STAT family members (Tang et al., 1994; O'Farrell et al., 1996; Jacobs-Helber et al., 1997; Joneja et al., 1997; Pearson et al., 1998). It remains unclear if these differences relate to technical parameters or possibly to lineage-specific differences in cell lines. Another family of signaling components activated in response to SCF are Src family members. SCF induces association of Lyn with phosphorylated tyrosine residues in the juxtamembrane region of the SCFR, as well as increases in both Lyn tyrosine phosphorylation and kinase activity (Linnekin et al., 1997b). Inhibition of Lyn activity with PP1, a Src-family inhibitor, or reduction in Lyn protein with antisense oligonucleotides, inhibits SCF-induced proliferation. In cells lacking Lyn, SCF activates other Src family members. Further, SCF induces increases in Fyn activity in mast cells transfected with murine SCFR. In vitro, Fyn binds a phosphopeptide sequence spanning tyrosines 568 and 570 of human SCFR, while

interaction of murine SCFR with the SH2 domain of Fyn requires tyrosine 567 (Price et al., 1997; Timokhina et al., 1998). Expression of Y567F murine SCF in SCFR-deficient murine mast cells inhibits activation of Fyn in response to SCF as well as partially inhibiting SCF-induced proliferation (Timokhina et al., 1998). Tec, a Src-like kinase related to Btk and Akt, is constitutively associated with the SCFR (Tang et al., 1994). Stimulation with SCF also results in increases in tyrosine phosphorylation of Tec, as well as increases in its catalytic activity. The role of Tec in SCF-mediated responses is presently unknown. The Ras/Raf/MAP kinase cascade is a signaling pathway activated in response to many growth factors (Madhani et al., 1998). SCF induces association of the SCFR with Grb2, Grap, SHP2, and Shc (Cutler et al., 1993; Matsuguchi et al., 1994; Tauchi et al., 1994a, 1994b; Feng et al., 1996). This in turn leads to colocalization of Sos and Ras and subsequent activation of Ras. RasGAP, Nf1, SHIP, and Vav may also play a role in modulating Ras activity in response to SCF (Miyazawa et al., 1991; Alai et al., 1992; Matsuguchi et al., 1995; Liu et al., 1997; Zhang et al., 1998). Activation of Ras results in activation of c-Raf-1, a serine threonine kinase critical in SCFmediated responses (Brennscheidt et al., 1994; Muszynski et al., 1995). SCF also induces increases in tyrosine phosphorylation and kinase activity of MEK1, MEK2, Erk1, and Erk2 (Hallek et al., 1992; Okuda et al., 1992; Tsai et al., 1993; Jacobs-Helber et al., 1997; Joneja et al., 1997; Pearson et al., 1998; Suie et al., 1998). Downstream of the Erks, the S6 kinase pp90rsk is activated (Tsai et al., 1993). In melanocytes, one substrate of the Erks is the microphthalmia transcription factor (MITF). Serine phosphorylation of MITF results in interaction with the transcriptional coactivator p300/CBP and transactivation (Hemesath et al., 1998; Price et al., 1998). In addition to its effects on the classical MAP kinase pathway, SCF also activates JNK, a MAP kinase involved in stress responses (Folz and Schrader, 1997). Recent studies by Besmer and coworkers indicate that activation of JNK is mediated by convergence of PI-3 kinase and Src signaling pathways on the GTPase Rac1 (Timokhina et al., 1998). SCF induces tyrosine phosphorylation of phospholipase C (PLC ), as well as a weak association between the SH2 domain of PLC and tyrosine 936 of the human SCFR (Lev et al., 1991; Reith et al., 1991; Rottapel et al., 1991; Hallek et al., 1992; BlumeJensen et al., 1994; Herbst et al., 1995b). The truncated form of the SCFR expressed in spermatids contains tyrosine 936. Interestingly, injection of mouse oocytes with this form of the SCFR activates

1922 Diana Linnekin and Jonathan R. Keller PLC in oocytes and induces parthenogenesis (Sette et al., 1997, 1998). Although PLC is a substrate for the SCFR, it is unlikely that SCF activates the classical PLC pathway. SCF does not induce increases in inositol trisphosphate (IP3) levels and increases in diacylglycerol (DAG) are likely mediated by phospholipase D (Lev et al., 1991; Koike et al., 1993; Kozawa et al., 1997). Possible negative regulators of SCFR signaling are SHP1, SOCS, SHIP, Chk, and PKC isoforms (BlumeJenson et al., 1993, 1994, 1995; Lorenz et al., 1996; Paulson et al., 1996; Grgurevich et al., 1997; Huber et al., 1998; Kozlowski et al., 1998; De Sepulveda et al., 1999). SHP2 may also be a negative regulator of SCFR signaling in more differentiated lineages (Kozlowski et al., 1998). Figure 3 summarizes which pathways are likely targeted by these signaling components. In summary, SCF activates multiple signal transduction components, including PI-3 kinase, Src family members, the Ras/Raf/MAP kinase cascade and the JAK/STAT pathway (Figure 2 and Figure 3). Activation of many of these pathways is dependent on interaction of an upstream signaling component with specific sites on the SCFR. Table 3 summarizes sites on the SCFR that interact with these signaling components as well as some of the known structurefunction relationships.

F

Shp1

PKC Src family members

SC

SCF

Figure 3 Summary of putative negative regulators of SCF-mediated signaling pathways. Src family members are negatively regulated by Chk, PKC isoforms, and SHP1. SOCS and SHP1 probably regulate SCF-induced activation of JAK family members. The Ras/Raf/MAP kinase cascade can be regulated by SHIP1, SHP1, SHP2, and PKC isoforms, while PI-3 kinase activation can be regulated by PKC isoforms and SHIP.

ChK Shp1

PKC Ras-Raf-Map kinase cascade Shp2 Ship

Socs Jak/Stat pathway

PI3 kinase

Downstream of the signaling pathways described above, pathways involved in mediating cell survival and cell cycle progression are activated in response to SCF. Although SCF prevents apoptosis in hematopoietic cells, in some epithelial cells it may be proapoptotic. Thus, the effects of SCF on cell survival are dependent on cellular context. With regards to the antiapoptotic effects of SCF, PI-3 kinase clearly plays a role. One kinase downstream of PI-3 kinase and involved in SCF-mediated survival is Akt. SCFinduced activation of Akt in fibroblasts expressing wild-type human SCFR resulted in interaction of Akt and Bad, and serine phosphorylation of Bad (BlumeJensen et al., 1998). SCF inhibits p53-mediated apoptosis in erythroleukemia cell lines (Abrahamson et al., 1995; Lin and Benchimol, 1995; Lee, 1998). Further, there is a reduced rate of apoptosis in SCF-deprived, bone marrow-derived mast cells from p53-deficient mice as compared with animals expressing normal levels of p53 (Yee et al., 1994b). Signaling pathways involved in SCF-induced cell cycle progression have also been examined in recent years. SCF induces events classically associated with the G1 /S transition, such as increases in Cdk2 activity and phosphorylation of Rb (Mantel et al., 1996). SCF also induces increases in expression of CaM-BP68, a calmodulin-binding protein likely involved in the G1 / S transition of the cell cycle (Reddy and Quesenberry, 1996). One group has suggested that synergistic responses induced by SCF in combination with GMCSF are mediated by alterations in expression of the Cdk inhibitors p21cip-1 and p27kip-1 (Mantel et al., 1996).

DOWNSTREAM GENE ACTIVATION

Transcription factors activated SCF induces increases in both the expression and activity of a variety of transcription factors. Several groups have reported SCF-induced increases in tyrosine phosphorylation and DNA-binding activity of STAT family members, including STAT1 and STAT5 (DeBerry et al., 1997; Ryan et al., 1997). SCF also induces increases in serine phosphorylation of STAT3 (Gotoh et al., 1996). As discussed above, one transcription factor studied rather extensively in relation to SCF signaling is MITF. SCF induces serine phosphorylation of MITF and interaction with the coactivator p300/ CBP. This complex binds DNA and activates DNA transcription (Hemesath et al., 1998; Price et al., 1998).

SCF Receptor 1923 Table 3 Signaling components that associate with specific regions of the SCFR and potential function Protein

Region

Tyrosine residue(s) (human/murine sequence)

Function

Cell line

Reference

Lyn

Juxta

Within 544±577 (human)

Proliferation

Mo7e

Linnekin et al., 1997b

Fyn

Juxta

Y568, Y570 (human)

±

±

Price et al., 1997

Fyn

Juxta

Y567 (murine)

Partial effect survival

Mast cells

Timokhina et al., 1998

Fyn

Juxta

Y567 (murine)

Partial effect proliferation

Mast cells

Timokhina et al., 1998

Shc

Juxta

Y568, Y570 (human)

±

±

Price et al., 1997

SHP1

Juxta

Y569 (murine)

Negative regulator of proliferation

BaF3

Kozlowski et al., 1998

SHP2

Juxta

Y567 (murine)

Negative regulator of proliferation

BaF3

Kozlowski et al., 1998

Chk

Juxta

Y568, Y570 (human)

±

±

Price et al., 1997

p85 P-I3 kinase

Insert

Y719 (murine)

Adhesion

Mast cells

Serve et al., 1995

Insert

Y719 (murine)

Partial effect survival, proliferation

Mast cells

Serve et al., 1995

Insert

Y719 (murine)

c-Kit trafficking

DA1 cells

Gommerman et al., 1997

Insert

Y721 (human)

Differentiation

FDC-P1 cells

Kubota et al., 1998

Insert

Y721 (human)

Survival

Fibroblasts

Blume-Jenson et al., 1998

Kinase II

Y821 (murine)

Survival

Mast cells

Serve et al., 1995

Kinase II

Y821 (murine)

Proliferation

Mast cells

Serve et al., 1995

Tail

Y936 (human)

Parthenogenesis

Oocytes

Herbst et al., 1995b; Sette et al., 1997

Unknown PLC

Transcription factors induced by SCF include a number of immediate early-response genes. SCF induces increases in expression of c-fos, junB, egr-1 and c-myc (Horie and Broxmeyer, 1993; Serve et al., 1995; O'Farrell et al., 1996). Further, synergistic induction of these genes may play a role in the capacity of SCF to synergize with other growth factors (Horie and Broxmeyer, 1993). SCF also induces increases in expression of SCL and c-Myb. As described below, both of these transcription factors play a role in maintaining expression of the SCFR (Ratajczak et al., 1992; Miller et al., 1994; Melotti and Calabretta, 1996; Krosl et al., 1998).

Genes induced Several classes of signaling molecules are induced in response to SCF. Csk-homologous kinase (Chk) is a kinase involved in downregulation of Src family kinase activity. SCF induces increases in Chk expression that peak 8 hours after stimulation and are maintained for 24 hours (Grgruvevich et al.,

1997). Studies with phosphopeptides suggest that, similar to Lyn, Chk associates with phosphorylated tyrosine residues in the juxtamembrane region of the SCFR (Price et al., 1997). Similarly, SOCS, an inhibitor of JAK family tyrosine kinases, is also induced in response to SCF (De Sepulveda et al., 1999). Another signaling molecule induced after stimulation of Mo7e cells with SCF in combination with IFN is the serine/threonine kinase Pim1 (YipSchneider et al., 1995). Pim1 was also induced by IL-3 and SCF treatment of the mast cell line B6M (O'Farrell et al., 1996). In contrast, in FDC2 cells, SCF as a single factor induced expression of Pim1 (Joneja et al., 1997). Possible explanations for the differences in these findings could be differences in either cell lineage or the differentiation state of these cell lines. The role of Pim1 in responses mediated by SCF alone or in combination with other factors remains to be defined. Recently, Keller and coworkers found that SCF in combination with IL-3 induces D3 in lineage-negative hematopoietic progenitor cells (Weiler et al., 1999). Although the function of this protein is not yet

1924 Diana Linnekin and Jonathan R. Keller known, expression correlates with myeloid differentiation in both cell lines and normal progenitor cells. Lastly, SCF also induces a number of immediate early-response genes such as c-fos, junB, egr-1, and c-myc (Horie and Broxmeyer, 1993; Serve et al., 1995; O'Farrell et al., 1996).

BIOLOGICAL CONSEQUENCES OF ACTIVATING OR INHIBITING RECEPTOR AND PATHOPHYSIOLOGY

Promoter regions involved

Unique biological effects of activating the receptors

The promoter region for both the murine and human SCFR has been cloned and partially characterized. Analysis of the sequence suggests multiple transcription factors regulate expression of the SCFR in different tissues. Included among these are AP-2, SP-1, Myb/Ets-family members and SCL. Early studies of the SCFR promoter were reviewed in 1994 by Galli and coworkers. This section will review recent data relating to regulation of the SCFR promoter. One transcription factor regulating SCFR expression in mast cells and melanocytes is MITF, a member of the basic-helix-loop-helix-leucine zipper family (bHLH-Zip) Similar to white spotting mice, mice deficient in MITF have reduced numbers of mast cells, as well as defects in pigmentation. Interestingly, the SCFR is not expressed in either cultured mast cells or melanocytes from these mice, but is expressed at normal levels in erythroid progenitors and germ cells (Isozaki et al., 1994). These findings suggest tissue-specific elements are involved in MITF regulation of SCFR expression. While the SCFR promoter has three CANNTG motifs that could serve as binding sites for bHLH-Zip transcription factors, only one binds MITF (Tsujimura et al., 1996a). Another bHLH-Zip transcription factor with potential importance in the regulation of SCFR expression is SCL, also known as TAL1. Inhibition of SCL expression by transfecting cells with SCL cDNA in the antisense orientation reduces expression of SCFR protein (Krosl et al., 1998). Similar results are obtained when cells are transfected with a dominant negative SCL construct. The induction of SCL by SCF during erythroid differentiation suggests a positive feedback loop for SCFR expression (Miller et al., 1994). The c-Kit promoter also contains multiple AP-2 binding sites (Giebel et al., 1992; Yamamoto et al., 1993; Yasuda et al., 1993). AP-2 binds the SCFR promoter and upregulates SCFR expression. Downregulation of AP-2 in melanoma cells results in downregulation of the SCFR and may contribute to an increase in metastatic potential (Huang et al., 1998). Other transcription factors with binding sites in the c-Kit promoter region are SP-1, Myb, and Ets (Ratajczak et al., 1992; Melloti and Calabretta, 1996; Park et al., 1998; Ratajczak et al., 1998).

The transforming capabilities of the SCFR are illustrated by the oncogenic potential of v-Kit. Structurally, v-Kit lacks the extracellular region and the 50 amino acid C-terminus of the SCFR (Qiu et al., 1988; Herbst et al., 1995a). In addition, there is a five amino acid insertion in the carboxyl tail, a deletion of two amino acids in the juxtamembrane region and one point mutation in the kinase insert region. As expected, v-Kit has constitutive tyrosine kinase activity in vitro (Majumder et al., 1990). Other constitutively active forms of the SCFR have been identified and are associated with cellular transformation. One of the best characterized gainof-function mutations in the human SCFR is substitution of valine for aspartic acid 816 (aspartic acid 814 of murine SCFR). This mutant has been identified in factor-independent mastocytoma lines derived from humans (HMC1), mice (P-815) and rats (RBL2H3) (Furitsu et al., 1993; Tsujimura et al., 1994). Expression of D816V SCFR in factor-dependent cell lines results in factor-independent cell lines that produce tumors in mice (Kitayama et al., 1995, 1996; Piao and Bernstein, 1996; Ferrao et al., 1997). As discussed above, this mutation is also found in patients with mastocytosis with associated hematological disorders, as well as in patients with other forms of mastocytosis (Nagata et al., 1995; Longley et al., 1996, 1999; Afonja et al., 1998; BuÈttner et al., 1998; Worobec et al., 1998). The D816V SCFR mutant is constitutively phosphorylated on tyrosine residues and the catalytic activity is increased as compared with wild-type SCFR (Furitsu et al., 1993; Tsujimura et al., 1994; Kitayama et al., 1995; Piao and Bernstein, 1996; Piao et al., 1996). D816V SCFR may also have alterations in substrate specificity that result in autophosphorylation of different sites (Piao et al., 1996). In addition, accelerated degradation of SHP1, a negative regulator SCFR signaling, has also been reported. Although D816V SCFR is constitutively active, it does not form dimers through the extracellular domain (Kitayama et al., 1995). In contrast, this form of the SCFR may dimerize through regions of the intracellular domain (Lam et al., 1999; Tsujimura et al., 1999).

SCF Receptor 1925 Several gain-of-function mutations of the SCFR have been detected in the juxtamembrane region. Substitution of glycine for valine 559 of murine SCFR results in constitutive dimerization, activation, and increases in oncogenic potential (Kitayama et al., 1995, 1996; Tsujimura, 1996; Tsujimura et al., 1997). Deletion of seven amino acids in the juxtamembrane region also generates a constitutively active form of the SCFR that spontaneously dimerizes (Tsujimura et al., 1996b). In addition, a series of mutations in the SCFR juxtamembrane region resulting in constitutive kinase activity have been found in some patients with gastrointestinal stromal tumors, as well as dogs with mastocytosis (Hirota et al., 1998; Ma et al., 1999; Moskaluk et al., 1999). Table 1 summarizes activating mutations found in the SCFR and diseases with which they are associated.

Phenotypes of receptor knockouts and receptor overexpression mice Studies of mice with a variety of mutations in the white spotting locus have demonstrated the importance of the SCFR in hematopoiesis, melanocyte, and germ cell development, and in intestine and brain function (Russell, 1979; Galli et al., 1994; Huizinga et al., 1995; Motro et al., 1996). The molecular basis of the known white spotting mutants and the corresponding phenotypes have previously been described (Nocka et al., 1990; Reith et al., 1991; Fleischman, 1993; Morrison-Graham and Takahashu, 1993). In summary, the animals die in utero at mid-gestation. In addition, heterozygotic animals expressing kinase-defective SCFR also have severe phenotypes. This dominant negative effect is the result of dimerization of wild-type and kinase inactive SCFR and subsequent impairment in signaling.

Human abnormalities Mutations in the SCFR have been detected in patients with autosomal dominant piebaldism. These individuals have aberrations in pigmentation on the abdomen, extremities, and forehead. Interestingly, the distribution of the pigmentation defects is similar to that of the white spotting mice. The mutations in several patients have been identified and occur predominantly in the SCFR catalytic domain. Alterations in amino acids 561, 583, 584, 642, and 664 have all been described (Spritz, 1994). Although no abnormalities in hematopoiesis have been reported, all of the

patients examined were heterozygotes (Spritz, 1992). Many of the heterozygotic white spotting mice reported to date also have normal hematopoiesis. As discussed previously, activated forms of the SCFR have been found in patients with gastrointestinal stromal tumors and mastocytosis (Nagata et al., 1995; Longley et al., 1996, 1999; Hirota et al., 1998; Moskaluk et al., 1999).

THERAPEUTIC UTILITY

Effect of treatment with soluble receptor domain As discussed above, SCF binds the SCFR extracellular domain and induces dimerization. The soluble form of the SCFR binds SCF and antagonizes SCF-induced increases in autophosphorylation as well as activation of downstream signaling pathways (Lev et al., 1992a). This results in inhibition of SCFmediated responses (Lev et al., 1992a, 1993). To date this has not been utilized clinically.

Effects of inhibitors (antibodies) to receptors AG1296 is a quinoxaline tyrphostin compound that inhibits the kinase activity of the PDGF and SCF receptors (Kovalenko et al., 1994). Because a high percentage of SCLC cells inappropriately express both SCF and its receptor, Krystal and coworkers examined the effect of AG1296 on the growth of SCLC cell lines in vitro (Krystal et al., 1997). This drug inhibited SCLC growth as well as induced apoptosis. These results suggest that an autocrine loop mediates survival and growth of some SCLC cells. Another drug that is reported to inhibit SCFRmediated signaling is AG776 (Wessely et al., 1997). Treatment of avian erythroid progenitor cells with this drug decreased SCF-mediated survival (Wessely et al., 1997). To date, there are no published reports relating to the use of either of these drugs in patients. In humans, antibodies blocking the binding of SCF to its receptor have not been used in vivo. However, recent studies in vitro suggest that the SCFR may be utilized to transfer genes into hematopoietic progenitor cells. Keller and coworkers demonstrated targeted transfection of hematopoietic cell lines expressing the SCFR with a molecular conjugate vector containing SCF (Schwarzenberger et al., 1996). SCF has also been incorporated into retroviral vectors and utilized

1926 Diana Linnekin and Jonathan R. Keller successfully for targeted gene transfer in vitro (Yajima et al., 1998). Lastly, a plasmid containing IL-3 was recently conjugated to an antibody specific for the SCFR and transiently transfected into a hematopoietic cell line as well as CD34-positive progenitor cells (Chapel et al., 1999). Thus, the SCFR may be an ideal target for gene transfer into hematopoietic cells.

References Abkowitz, J. L., Broudy, V. C., Bennett, L. G., Zsebo, K. M., and Martin, F. H. (1992). Absence of abnormalities of c-kit or its ligand in two patients with diamond-blackfan anemia. Blood 79, 25±28. Abrahamson, J. L. A., Lee, J. M., and Bernstein, A. (1995). Regulation of p53-mediated apoptosis and cell cycle arrest by Steel factor. Mol. Cell. Biol. 15, 6953±6960. Afonja, O., Amorosi, E., Ashman, L., and Takeshita, K. (1998). Multilineage involvement and erythropoietin-`independent' erythroid progenitor cells in a patient with systemic mastocytosis. Ann. Hematol. 77, 183±186. Alai, M., Mui, A. L. -F., Cutler, R. L., Bustelo, S. R., Barbacid, M., and Krystal, G. (1992). Steel factor stimulates the tyrosine phosphorylation of the proto-oncogene product, p95vav , in human hemopoietic cells. J. Biol. Chem. 267, 18021±18025. AndreÂ, C., Martin, E., Cornu, F., Hu, W.-X., Wang, X.-P., and Galibert, F. (1992). Genomic organization of the human c-kit gene: evolution of the receptor tyrosine kinase subclass III. Oncogene 7, 685±691. Asano, Y., Brach, M. A., Ahlers, A., de Vos, S., Butterfield, J. H., Ashman, L. K., Valent, P., Gruss, H.-J., and Herrmann, F. (1993). Phorbol ester 12-O-tetradecanoylphorbol-13-acetate down-regulates expression of the c-kit proto-oncogene product. J. Immunol. 151, 2345±2354. Ashman, L. (1999). The biology of stem cell factor and its receptor c-Kit. Int. J. Biochem. Cell Biol. 31, 1037±1051. Badache, A., Muja, N., and De Vries, G. H. (1998). Expression of Kit in neurofibromin-deficient human Schwann cells: role in Schwann cell hyperplasia associated with type 1 neurofibromatosis. Oncogene 17, 795±800. Besmer, P., Murphy, J. E., George, P. C., Qiu, F., Bergold, P. J., Lederman, L., Snyder Jr. H. W., Brodeur, D., Zuckerman, E. E., and Hardy, W. D. (1986). A new acute transforming feline retrovirus and relationship of its oncogene v-kit with the protein kinase gene family. Nature 320, 415±421. Blechman, J. M., Lev, S., Brizzi, M. F., Leitner, O., Pegoraro, L., Givol, D., and Yarden, Y. (1993a). Soluble c-kit proteins and antireceptor monoclonal antibodies confine the binding site of the stem cell factor. J. Biol. Chem. 268, 4399±4406. Blechman, J. M., Lev, S., Givol, D., and Yarden, Y. (1993b). Structure±function analyses of the kit receptor for the steel factor. Stem Cells 11, 12±21. Blechman, J. M., Lev, S., Barg, J., Eisenstein, M., Vaks, B., Vogel, Z., Givol, D., and Yarden, Y. (1995). The fourth immunoglobulin domain of the stem cell factor receptor couples ligand binding to signal transduction. Cell 80, 103±113. Blume-Jensen, P., Claesson-Welsh, L., Siegbahn, A., Zsebo, K. M., Westermark, B., and Heldin, C.-H. (1991). Activation of the human c-kit product by ligand-induced dimerization mediates circular actin reorganization and chemotaxis. EMBO J. 10, 4121±4128.

Blume-Jensen, P., Siegbahn, A., Stabel, S., Heldin, C.-H., and RoÈnnstrand, L. (1993). Increased Kit/SCF receptor induced mitogenicity but abolished cell motility after inhibition of protein kinase C. EMBO J. 12, 4199±4209. Blume-Jensen, P., RoÈnnstrand, L., Gout, I., Waterfield, M. D., and Heldin, C.-H. (1994). Modulation of Kit/stem cell factor receptor-induced signaling by protein kinase C. J. Biol. Chem. 269, 21793±21802. Blume-Jensen, P., Wernstedt, C., Heldin, C.-H., and RoÈnnstrand, L. (1995). Identification of the major phosphorylation sites for protein kinase C in Kit/stem cell factor receptor in vitro and in intact cells. J. Biol. Chem. 270, 14192±14200. Blume-Jensen, P., Janknecht, R., and Hunter, T. (1998). The Kit receptor promotes cell survival via activation of PI 3-kinase and subsequent Akt-mediated phosphorylation of Bad on Ser136. Curr. Biol. 8, 779±782. Boissy, R. E., and Nordlund, J. J. (1997). Molecular basis of congenital hypopigmentary disorders in humans: a review. Pigment Cell Res. 10, 12±24. Bondy, C. A., Nelson, L. M., and Kalantaridou, S. N. (1998). The genetic origins of ovarian failure. J. Women's Health 7, 1225±1229. BosseÂ, P., Bernex, F., De Sepulveda, P., SalauÈn, P., Panthier, J.-J. (1997). Multiple neuroendocrine tumours in transgenic mice induced by c-kit-SV40 T antigen fusion genes. Oncogene 14, 2661±2670. Brennscheidt, U., Riedel, D., KoÈlch, W., Bonifer, R., Brach, M. A., Ahlers, A., Mertelsmann, R. H., and Herrmann, F. (1994). Raf-1 is a necessary component of the mitogenic response of the human megakaryoblastic leukemia cell line MO7 to human stem cell factor, granulocyte-macrophage colony-stimulating factor, interleukin 3, and interleukin 9. Cell Growth Differ. 5, 367±372. Brizzi, M. F., Blechman, J. M., Cavalloni, G., Givol, D., Yarden, Y., and Pegoraro, L. (1994a). Protein kinase C-dependent release of a functional whole extracellular domain of the mast cell growth factor (MGF) receptor by MGF-dependent human myeloid cells. Oncogene 9, 1583±1589. Brizzi, M. F., Zini, M. G., Aronica, M. G., Blechman, J. M., Yarden, Y., and Pegoraro, L. (1994b). Convergence of signaling by interleukin-3, granulocyte-macrophage colony-stimulating factor, and mast cell growth factor on JAK2 tyrosine kinase. J. Biol. Chem. 269, 31680±31684. Broudy, V. C. (1997). Stem cell factor and hematopoiesis. Blood 90, 1345±1364. Broudy, V. C., Lin, N. L., BuÈhring, H.-J., Komatsu, N., and Kavanagh, T. J. (1998). Analysis of c-kit receptor dimerization by fluorescence resonance energy transfer. Blood 91, 898±906. BuÈttner, C., Henz, B. M., Welker, P., Sepp, N. T., and Grabbe, J. (1998). Identification of activating c-kit mutations in adult-, but not in childhood-onset indolent mastocytosis: a possible explanation for divergent clinical behavior. J. Invest. Dermatol. 111, 1227±1231. Carpino, N., Wisniewski, D., Strife, A., Marshak, D., Kobayashi, R., Stillman, B., and Clarkson, B. (1997). p62dok : a constitutively tyrosine-phosphorylated, GAP-associated protein in chronic myelogenous leukemia progenitor cells. Cell 88, 197±204. Chabot, B., Stephenson, D. A., Chapman, V. M., Besmer, P., and Bernstein, A. (1988). The proto-oncogene c-kit encoding a transmembrane tyrosine kinase receptor maps to the mouse W locus. Nature 335, 88±89. Chan, J. K. (1999). Mesenchymal tumors of the gastrointestinal tract: a paradise for acronyms (STUMP, GIST, GANT, and now GIPACT), implication of c-kit in genesis, and yet another

SCF Receptor 1927 of the many emerging roles of the interstitial cell of Cajal in the pathogenesis of gastrointestinal diseases? Adv. Anat. Pathol. 6, 19±40. Chapel, A., Poncet, P., Neildez-Nguyen, T. M. A., VeÂtillard, J., Brouard, N., Goupy, C., Chavanel, G., Hirsch, F., and Thierry, D. (1999). Targeted transfection of the IL-3 gene into primary human hematopoietic progenitor cells through the c-kit receptor. Exp. Hematol. 27, 250±258. Chui, X., Egami, H., Yamashita, J., Kurizaki, T., Ohmachi, H., Yamamoto, S., and Ogawa, M. (1996). Immunohistochemical expression of the c-kit proto-oncogene product in human malignant and non-malignant breast tissues. Br. J. Cancer 73, 1233±1236. Cutler, R. L., Liu, L., Damen, J. E., and Krystal, G. (1993). Multiple cytokines induce the tyrosine phosphorylation of Shc and its association with Grb2 in hemopoietic cells. J. Biol. Chem. 268, 21463±21465. DeBerry, C., Mou, S., and Linnekin, D. (1997). Stat1 associates with c-kit and is activated in response to stem cell factor. Biochem. J. 327, 73±80. De Sepulveda, P., Okkenhaug, K., La Rose, J., Hawley, R. G., Dubreuil, P., and Rottapel, R. (1999). Socs1 binds to multiple signalling proteins and suppresses Steel factor-dependent proliferation. EMBO J. 18, 904±915. de Vos, S., Brach, M. A., Asano, Y., Ludwig, W.-D., Beffelheim, P., Gruss, H.-J., and Herrmann, F. (1993). Transforming growth factor- 1 interferes with the proliferation-inducing activity of stem cell factor in myelogenous leukemia blasts through functional down-regulation of the c-kit proto-oncogene product. Cancer Res. 53, 3638±3642. DiPaola, R. S., Kuczynski, W. I., Onodera, K., Ratajczak, M. Z., Hijiya, N., Moore, J., and Gewirtz, A. M. (1997). Evidence for a functional kit receptor in melanoma, breast, and lung carcinoma cells. Cancer Gene Ther. 4, 176±182. Donovan, P. J. (1994). Growth factor regulation of mouse primordial germ cell development. Curr. Topics Dev. Biol. 29, 189±225. Donovan, P. J., De Miguel, M., Cheng, L., and Resnick, J. L. (1998). Primordial germ cells, stem cells and testicular cancer. APMIS 106, 134±141. Driancourt, M.-A., and Thuel, B. (1998). Control of oocyte growth and maturation by follicular cells and molecules present in follicular fluid. Reprod. Nutr. Dev. 38, 345±362. Dubois, C. M., Ruscetti, F. W., Stankova, J., and Keller, J. R. (1994). Transforming growth factor-beta regulates c-kit message stability and cell-surface protein expression in hematopoietic progenitors. Blood 83, 3138±3145. Escribano, L., Ocqueteau, M., Almeida, J., Orfao, A., and San Miguel, J. F. (1998). Expression of the c-kit (CD117) molecule in normal and malignant hematopoiesis. Leuk. Lymph. 30, 459±466. Feng, G.-S., Ouyang, Y.-B., Hu, D.-P., Shi, Z-Q., Gentz, R., and Ni, J. (1996). Grap is a novel SH3-SH2-SH3 adaptor protein that couples tyrosine kinases to the ras pathway. J. Biol. Chem. 271, 12129±12132. Feng, H., Sandlow, J. I., and Sandra, A. (1998). The C-kit receptor and its possible signaling transduction pathway in mouse spermatozoa. Mol. Reprod. Dev. 49, 317±326. Ferrao, P., Gonda, T. J., and Ashman, L. K. (1997). Expression of constitutively activated human c-kit in myb transformed early myeloid cells leads to factor independence, histiocytic differentiation, and tumorigenicity. Blood 90, 4539±4552. Fleischman, R. A. (1993). From white spots to stem cells: the role of the Kit receptor in mammalian development. Trends Genet. 9, 285±290.

Fleischman, R. A., Saltman, D. L., Stastny, V., and Zneimer, S. (1991). Deletion of the c-kit protooncogene in the human developmental defect piebald trait. Proc. Natl Acad. Sci. USA 88, 10885±10889. FoÈdinger, M., and Mannhalter, C. (1997). Molecular genetics and development of mast cells: implications for molecular medicine. Mol. Med. Today 3, 131±137. Foltz, I. N., and Schrader, J. W. (1997). Activation of the stressactivated protein kinases by multiple hematopoietic growth factors with the exception of interleukin-4. Blood 89, 3092±3096. Fujio, K., Hu, Z., Evarts, R. P., Marsden, E. R., Niu, C.-H., and Thorgeirsson, S. S. (1996). Coexpression of stem cell factor and c-kit in embryonic and adult liver. Exp. Cell Res. 224, 243±250. Funasaka, Y., Boulton, T., Cobb, M., Yarden, Y., Fan, B., Lyman, S. D., Williams, D. E., Anderson, D. M., Zakut, R., Mishima, Y., and Halaban, R. (1992). c-Kit-kinase induces a cascade of protein tyrosine phosphorylation in normal human melanocytes in response to mast cell growth factor and stimulates mitogen-activated protein kinase but is down-regulated in melanomas. Mol. Biol. Cell 3, 197±209. Furitsu, T., Tsujimura, T., Tono, T., Ikeda, H., Kitayama, H., Koshimizu, U., Sugahara, H., Butterfield, J. H., Ashman, L. K., Kanayama, Y., Matsuzawa, Y., Kitamura, Y., and Kanakura, Y. (1993). Identification of mutations in the coding sequence of the proto-oncogene c-kit in a human mast cell leukemia cell line causing ligand-independent activation of c-kit product. J. Clin. Invest. 92, 1736±1744. Galli, S. J., Zsebo, K. M., and Geissler, E. N. (1994). The kit ligand, stem cell factor. Adv. Immunol. 55, 1±97. Geissler, E. N., Ryan, M. A., and Housman, D. E. (1988). The dominant-white spotting (W) locus of the mouse encodes the c-kit proto-oncogene. Cell 55, 185±192. Giebel, L. B., and Spritz, R. A. (1991). Mutation of the KIT (mast/ stem cell growth factor receptor) protooncogene in human piebaldism. Proc. Natl Acad. Sci. USA 88, 8696±8699. Giebel, L. B., Strunk, K. M., Holmes, S. A., and Spritz, R. A. (1992). Organization and nucleotide sequence of the human KIT (mast/stem cell growth factor receptor) proto-oncogene. Oncogene 7, 2207±2217. Gokkel, E., Grossman, Z., Ramot, B., Yarden, Y., Rechavi, G., and Givol, D. (1992). Structural organization of the murine c-kit proto-oncogene. Oncogene 7, 1423±1429. Gommerman, J. L., Rottapel, R., and Berger, S. A. (1997). Phosphatidylinositol 3-kinase and Ca2‡ influx dependence for ligand-stimulated internalization of the c-Kit receptor. J. Biol. Chem. 272, 30519±30525. Gotoh, A., Takahira, H., Mantel, C., Litz-Jackson, S., Boswell, H. S., and Broxmeyer, H. E. (1996). Steel factor induces serine phosphorylation of Stat3 in human growth factor-dependent myeloid cell lines. Blood 88, 138±145. Grgurevich, S., Linnekin, D., Musso, T., Zhang, X., Modi, W., Varesio, L., Ruscetti, F. W., Ortaldo, J. R., and McVicar, D. W. (1997). The Csk-like proteins Lsk, Hyl, and Matk represent the same Csk homologous kinase (Chk) and are regulated by stem cell factor in the megakaryoblastic cell line M07e. Growth Factors 14, 103±115. Gutman, M., Singh, R. K., Radinsky, R., and Bar-Eli, M. (1994). Intertumoral heterogeneity of receptor-tyrosine kinases expression in human melanoma cell lines with different metastatic capabilities. Anticancer Res. 14, 1759±1766. Hallek, M., Druker, B., Lepisto, E. M., Wood, K. W., Ernst, T. J., and Griffin, J. D. (1992). Granulocyte-macrophage colonystimulating factor and steel factor induce phosphorylation of both unique and overlapping signal transduction intermediates

1928 Diana Linnekin and Jonathan R. Keller in a human factor-dependent hematopoietic cell line. J. Cell Physiol. 153, 176±186. Hallek, M., Danhauser-Riedl, S., Herbst, R., Warmuth, M., Winkler, A., Kolb, H.-J., Druker, B., Griffin, J. D., Emmerich, B., and Ullrich, A. (1996). Interaction of the receptor tyrosine kinase p145c-kit with the p210bcr=abl kinase in myeloid cells. Br. J. Haematol. 94, 5±16. Hamel, W., and Westphal, M. (1997). The road less travelled: c-kit and stem cell factor. J. Neuro-Oncol. 35, 327±333. Hassan, H. T., and Zander, A. (1996). Stem cell factor as a survival and growth factor in human normal and malignant hematopoiesis. Acta Haematol. 95, 257±262. He, J., DeCastro, C. M., Vandenbark, G. R., Busciglioi, J., and Gabuzda, D. (1997). Astrocyte apoptosis induced by HIV-1 transactivation of the c-kit protooncogene. Proc. Natl Acad. Sci. USA 94, 3954±3959. Heinrich, M. C., Dooley, D. C., and Keeble, W. W. (1995). Transforming growth factor 1 inhibits expression of the gene products for Steel factor and its receptor (c-kit). Blood 85, 1769±1780. Hemesath, T. J., Price, E. R., Takemoto, C., Badalian T., and Fisher, D. E. (1998). MAP kinase links the transcription factor microphthalmia to c-Kit signalling in melanocytes. Nature 391, 298±301. Herbst, R., Munemitsu, S., and Ullrich, A. (1995a). Oncogenic activation of v-kit involves deletion of a putative tyrosine-substrate interaction site. Oncogene 10, 369±379. Herbst, R., Shearman, M. S., Jallal, B., Schlessinger, J., and Ullrich, A. (1995b). Formation of signal transfer complexes between stem cell and platelet-derived growth factor receptors and SH2 domain proteins in vitro. Biochemistry 34, 5971±5979. Hibi, K., Takahashi, T., Sekido, Y., Ueda, R., Hida, T., Ariyoshi, Y., Takagi, H., and Takahashi, T. (1991). Coexpression of the stem cell factor and the c-kit genes in small-cell lung cancer. Oncogene 6, 2291±2296. Hines, S. J., Organ, C., Kornstein, M. J., and Krystal, G. W. (1995). Coexpression of the c-kit and stem cell factor genes in breast carcinomas. Cell Growth Differ. 6, 769±779. Hirota, S., Ito, A., Morii, E., Wanaka, A., Tohyama, M., Kitamura, Y., and Nomura, S. (1992). Localization of mRNA for c-kit receptor and its ligand in the brain of adult rats: an analysis using in situ hybridization histochemistry. Mol. Brain Res. 15, 47±54. Hirota, S., Isozaki, K., Moriyama, Y., Hashimoto, K., Nishida, T., Ishiguro, S., Kawano, K., Hanada, M., Kurata, A., Takeda, M., Tunio, G. M., Matsuzawa, Y., Kanakura, Y., Shinomura, Y., and Kitamura, Y. (1998). Gain-of-function mutations of c-kit in human gastrointestinal stromal tumors. Science 279, 577±580. Hofstra, R. M., Landsvater, R. M., Ceccherini, I., Stulp, R. P., Stelwagen, T., Luo, Y., Pasini, B., Hoppener, J. W., van Amstel, H. K., Romeo, G., Lips, C. J. M., and Buys, C.H. C. M. (1994). A mutation in the RET proto-oncogene associated with multiple endocrine neoplasia type 2B and sporadic medullary thyroid carcinoma. Nature 367, 375±376. Horie, M., and Broxmeyer, H. E. (1993). Involvement of immediate-early gene expression in the synergistic effects of steel factor in combination with granulocyte-macrophage colony-stimulating factor or interleukin-3 on proliferation of a human factordependent cell line. J. Biol. Chem. 268, 968±973. Hu, Z.-B., Ma, W., Uphoff, C. C., Quentmeier, H., and Drexler, H. G. (1994). C-kit expression in human megakaryoblastic leukemia cell lines. Blood 83, 2133±2144. Huang, E., Nocka, K., Beier, D. R., Chu, T.-Y., Buck, J., Lahm, H.-W., Wellner, D., Leder, P., and Besmer, P. (1990). The hematopoietic growth factor KL is encoded by the S1 locus

and is the ligand of the c-kit receptor, the gene product of the W locus. Cell 63, 225±233. Huang, S., Luca, M., Gutman, M., McConkey, D. J., Langley, K. E., Lyman, S. D., and Bar-Eli, M. (1996). Enforced c-KIT expression renders highly metastatic human melanoma cells susceptible to stem cell factor-induced apoptosis and inhibits their tumorigenic and metastatic potential. Oncogene 13, 2339±2347. Huang, S., Jean, D., Luca, M., Tainsky, M. A., and Bar-Eli, M. (1998). Loss of AP-2 results in downregulation of c-KIT and enhancement of melanoma tumorigenicity and metastasis. EMBO J. 17, 4358±4369. Huber, M., Helgason, C. D., Scheid, M. P., Duronio, V., Humphries, R. K., and Krystal, G. (1998). Targeted disruption of SHIP leads to steel factor-induced degranulation of mast cells. EMBO J. 17, 7311±7319. Huizinga, J. D., Thuneberg, L., KluÈppel, M., Malysz, J., Mikkelsen, H. B., and Bernstein, A. (1995). W/kit gene required for interstitial cells of Cajal and for intestinal pacemaker activity. Nature 373, 347±349. Inoue, M., Kyo, S., Fujita, M., Enomoto, T., and Kondoh, G. (1994). Coexpression of the c-kit receptor and the stem cell factor in gynecological tumors. Cancer Res. 54, 3049±3053. Ismail, R. S., Okawara, Y., Fryer, J. N., and Vanderhyden, B. C. (1996). Hormonal regulation of the ligand for c-kit in the rat ovary and its effects on spontaneous oocyte meiotic maturation. Mol. Reprod. Dev. 43, 458±469. Ismail, R. S., DubeÂ, M., and Vanderhyden, B. C. (1997). Hormonally regulated expression and alternative splicing of kit ligand may regulate kit-induced inhibition of meiosis in rat oocytes. Dev. Biol. 184, 333±342. Isozaki, K., Tsujimura, T., Nomura, S., Morii, E., Koshimizu, U., Nishimune, Y., and Kitamura, Y. (1994). Cell type-specific deficiency of c-kit gene expression in mutant mice of mi/mi genotype. Am. J. Pathol. 145, 827±836. Jacobs-Helber, S. M., Penta, K., Sun, Z., Lawson, A., and Sawyer, S. T. (1997). Distinct signaling from stem cell factor and erythropoietin in HCD57 cells. J. Biol. Chem. 272, 6850± 6853. Jacobsen, F. W., Dubois, C. M., Rusten, L. S., Veiby, O. P., and Jacobsen, S. E. (1995). Inhibition of stem cell factor-induced proliferation of primitive murine hematopoietic progenitor cells signaled through the 75-kilodalton tumor necrosis factor receptor. Regulation of c-kit and p53 expression. J. Immunol. 154, 3732±3741. Joneja, B., Chen, H.-C., Seshasayee, D., Wrentmore, A. L., and Wojchowski, D. M. (1997). Mechanisms of stem cell factor and erythropoietin proliferative co-signaling in FDC2-ER cells. Blood 90, 3533±3545. Kauma, S., Huff, T., Krystal, G., Ryan, J., Takacs, P., and Turner, T. (1996). The expression of stem cell factor and its receptor, c-kit in human endometrium and placental tissues during pregnancy. J. Clin. Endocrinol. Metab. 81, 1261±1266. Keshet, E., Lyman, S. D., Williams, D. E., Anderson, D. M., Jenkins, N. A., Copeland, N. G., and Parada, L. F. (1991). Embryonic RNA expression patterns of the c-kit receptor and its cognate ligand suggest multiple functional roles in mouse development. EMBO J. 10, 2425±2435. Khoury, E., Andre, C., Pontvert-Delucq, S., Drenou, B., Baillou, C., Guigon, M., Najman, A., and Lemoine, F. M. (1994). Tumor necrosis factor  (TNF) downregulates c-kit proto-oncogene product expression in normal and acute myeloid leukemia CD34‡ cells via p55 TNF receptors. Blood 84, 2506±2514.

SCF Receptor 1929 Kimura, A., Nakata, Y., Katoh, O., and Hyodo, H. (1997). c-kit point mutation in patients with myeloproliferative disorders. Leuk. Lymph. 25, 281±287. Kitamura, Y., Hirota, S., and Nishida, T. (1998). Molecular pathology of c-kit proto-oncogene and development of gastrointestinal stromal tumors. Ann. Chir. Gyn. 87, 282±286. Kitayama, H., Kanakura, Y., Furitsu, T., Tsujimura, T., Oritani, K., Ikeda, H., Sugahara, H., Mitsui, H., Kanayama, Y., Kitamura, Y., and Matsuzawa, Y. (1995). Constitutively activating mutations of c-kit receptor tyrosine kinase confer factor-independent growth and tumorigenicity of factor-dependent hematopoietic cell lines. Blood 85, 790±798. Kitayama, H., Tsujimura, T., Matsumura, I., Oritani, K., Ikeda, H., Ishikawa, J., Okabe, M., Suzuki, M., Yamamura, K., Matsuzawa, Y., Kitamura, Y., and Kanakura, Y. (1996). Neoplastic transformation of normal hematopoietic cells by constitutively activating mutations of c-kit receptor tyrosine kinase. Blood 88, 995±1004. Koike, T., Hirai, K., Morita, Y., and Nozawa, Y. (1993). Stem cell factor-induced signal transduction in rat mast cells. J. Immunol. 151, 359±366. Kondoh, G., Hayasaka, N., Li, Q., Nishimune, Y., and Hakura, A. (1995). An in vivo model for receptor tyrosine kinase autocrineparacrine activation: auto-stimulated KIT receptor acts as a tumor promoting factor in papillomavirus-induced tumorigenesis. Oncogene 10, 341±347. Kovalenko, M., Gazit, A., Bohmer, A., Rorsman, C., Ronnstrand, L., Heldin, C. H., Waltenberger, J., Bohmer, F. D., and Levitzki, A. (1994). Selective plateletderived growth factor receptor kinase blockers reverse sis-transformation. Cancer Res. 54, 6106±6114. Kozawa, O., Blume-Jensen, P., Heldin, C.-H., and RoÈnnstrand, L. (1997). Involvement of phosphatidylinositol 30 -kinase in stemcell-factor-induced phospholipase D activation and arachidonic acid release. Eur. J. Biochem. 248, 149±155. Kozlowski, M., Larose, L., Lee, F., Le, D. M., Rottapel, R., and Siminovitch, K. A. (1998). SHP-1 binds and negatively modulates the c-Kit receptor by interaction with tyrosine 569 in the c-Kit juxtamembrane domain. Mol. Cell. Biol. 18, 2089±2099. Krosl, G., He, G., Lefrancois, M., Charron, F., RomeÂo, P.-H., Jolecoeur, P., Kirsch, I. R., Nemer, M., and Hoang, T. (1998). Transcription factor SCL is required for c-kit expression and c-Kit function in hemopoietic cells. J. Exp. Med. 188, 439±450. Krystal, G. W., Hines, S. J., and Organ, C. P. (1996). Autocrine growth of small cell lung cancer mediated by coexpression of c-kit and stem cell factor. Cancer Res. 56, 370±376. Krystal, G. W., Carlson, P., and Litz, J. (1997). Induction of apoptosis and inhibition of small cell lung cancer growth by the quinoxalin tyrphostins. Cancer Res. 57, 2203±2208. Kubota, T., Hikona, H., Sasaki, E., and Sakurai, M. (1994). Sequence of a bovine c-kit proto-oncogene cDNA. Gene 141, 305±306. Kubota, Y., Angelotti, T., Niederfellner, G., Herbst, R., and Ullrich, A. (1998). Activation of phosphatidylinositol 3-kinase is necessary for differentiation of FDC-P1 cells following stimulation of type III receptor tyrosine kinases. Cell Growth Differ. 9, 247±256. Laitinen, M., Rutanen, E.-M., and Ritvos, O. (1995). Expression of c-kit ligand messenger ribonucleic acids in human ovaries and regulation of their steady state levels by gonadotropins in cultured granulosa-luteal cells. Endocrinology 136, 4407±4414. Lam, L. P. Y., Chow, R. Y. K., and Berger, S. A. (1999). A transforming mutation enhances the activity of the c-Kit soluble tyrosine kinase domain. Biochem. J. 338, 131±138.

Lammie, A., Drobnjak, M., Gerald, W., Saad, A., Cote, R., and Cordon-Cardo, C. (1994). Expression of c-kit and kit ligand proteins in normal human tissues. J. Histochem. Cytochem. 42, 1417±1425. Lassam, N., and Bickford, S. (1992). Loss of c-kit expression in cultured melanoma cells. Oncogene 7, 51±56. Lee, J. M. (1998). Inhibition of p53-dependent apoptosis by the KIT tyrosine kinase: regulation of mitochondrial permeability transition and reactive oxygen species generation. Oncogene 17, 1653±1662. Lemmon, M. A., Pinchasi, D., Zhou, M., Lax, I., and Schlessinger, J. (1997). Kit receptor dimerization is driven by bivalent binding of stem cell factor. J. Biol. Chem. 272, 6311± 6317. Lev, S., Givol, D., and Yarden, Y. (1991). A specific combination of substrates is involved in signal transduction by the kitencoded receptor. EMBO J. 10, 647±654. Lev, S., Yarden, Y., and Givol, D. (1992a). A recombinant ectodomain of the receptor for the stem cell factor (SCF) retains ligand-induced receptor dimerization and antagonizes SCFstimulated cellular responses. J. Biol. Chem. 267, 10866±10873. Lev, S., Yarden, Y., and Givol, D. (1992b). Dimerization and activation of the kit receptor by monovalent and bivalent binding of the stem cell factor. J. Biol. Chem. 267, 15970±15977. Lev, S., Blechman, J., Nishikawa, S.-I., Givol, D., and Yarden, Y. (1993). Interspecies molecular chimeras of kit help define the binding site of the stem cell factor. Mol. Cell. Biol. 13, 2224± 2234. Li, Q., Kondoh, G., Inafuku, S., Nishimune, Y., and Hukura, A. (1996). Abrogation of c-kit/Steel factor-dependent tumorigenesis by kinase defective mutants of the c-kit receptor: c-kit kinase defective mutants as candidate tools for cancer gene therapy. Cancer Res. 56, 4343±4346. Lin, Y., and Benchimol, S. (1995). Cytokines inhibit p53-mediated apoptosis but not p53-mediated G1 arrest. Mol. Cell. Biol. 15, 6045±6054. Linnekin, D. (1999). Early signaling pathways activated by c-Kit in hematopoietic cells. Int. J. Biochem. Cell Biol., 31, 1053±1074. Linnekin, D., Mou, S., Deberry, C. S., Weiler, S. R., Keller, J. R., Ruscetti, F. W., and Longo, D. L. (1997a). Stem cell factor, the JAK-STAT pathway and signal transduction. Leuk. Lymphoma. 27, 439±444. Linnekin, D., DeBerry, C. S., and Mou, S. (1997b). Lyn associates with the juxtamembrane region of c-Kit and is activated by stem cell factor in hematopoietic cell lines and normal progenitor cells. J. Biol. Chem. 272, 27450±27455. Liu, Y.-C., Kawagishi, M., Kameda, R., and Ohashi, H. (1993). Characterization of a fusion protein composed of the extracellular domain of c-kit and the Fc region of human IgG expressed in a baculovirus system. Biochem. Biophys. Res. Commun. 197, 1094±1102. Liu, L., Damen, J. E., Ware, M., Hughes, M., and Krystal, G. (1997). SHIP, a new player in cytokine-induced signalling. Leukemia 11, 181±184. Longley, B. J., Tyrrell, L., Lu, S.-Z., Ma, Y.-S., Langley, K., Ding, T., Duffy, T., Jacobs, P., Tang, L. H., and Modlin, I. (1996). Somatic c-KIT activating mutation in urticaria pigmentosa and aggressive mastocytosis: establishment of clonality in a human mast cell neoplasm. Nature Genet. 12, 312±314. Longley Jr., B. J., Metcalfe, D. D., Tharp, M., Wang, X., Tyrrell, L., Lu, S.-Z., Heitjan, D., and Ma, Y. (1999). Activating and dominant inactivating c-KIT catalytic domain mutations in distinct clinical forms of human mastocytosis. Proc. Natl Acad. Sci. USA. 96, 1609±1614.

1930 Diana Linnekin and Jonathan R. Keller Lorenz, U., Bergemann, A. D., Steinberg, H. N., Flanagan, J. G., Li, X., Galli, S. J., and Neel, B. G. (1996). Genetic analysis reveals cell type-specific regulation of receptor tyrosine kinase c-Kit by the protein tyrosine phosphatase SHP1. J. Exp. Med. 184, 1111±1126. Loveland, K. L., and Schlatt, S. (1997). Stem cell factor and c-kit in the mammalian testis: lessons originating from Mother Nature's gene knockouts. J. Endocrinol. 153, 337±344. Lu, C., and Kerbel, R. S. (1994). Cytokines, growth factors and the loss of negative growth controls in the progression of human cutaneous malignant melanoma. Curr. Opin. Oncol. 6, 212±220. Luca, M. R., and Bar-Eli, M. (1998). Molecular changes in human melanoma metastasis. Histol. Histopathol. 13, 1225±1231. Lyman, S. D., and Jacobsen, S. E. W. (1998). C-kit ligand and Flt3 ligand: stem/progenitor cell factors with overlapping yet distinct activities. Blood 91, 1101±1134. Ma, Y., Longley, B. J., Wang, X., Blount, J. L., Langley, K., and Caughey, G. H. (1999). Clustering of activating mutations in c-KIT's juxtamembrane coding region in canine mast cell neoplasms. J. Invest. Dermatol. 112, 165±170. Madhani, H. D., and Fink, G. R. (1998). The riddle of MAP kinase signaling specificity. TIG 14, 151±155. Majumder, S., Ray, P., and Besmer, P. (1990). Tyrosine protein kinase activity of the HZ4-feline sarcoma virus P80gag-kit -transforming protein. Oncogene Res. 5, 329±335. Mantel, C., Luo, Z., Canfield, J., Braun, S., Deng, C., and Broxmeyer, H. E. (1996). Involvement of p21cip-1 in the maintenance of stem/progenitor cells in vivo. Blood 88, 3710±3719. Marklund, S., Kijas, J., Rodriguez-Martinez, H., RoÈnnstrand, L., Funa, K., Moller, M., Lange, D., Edfors-Lilja, I., and Andersson, L. (1998). Molecular basis for the dominant white phenotype in the domestic pig. Genome Res. 8, 826±833. Matsuda, R., Takahashi, T., Nakamura, S., Sekido, Y., Nishida, K., Seto, M., Seito, T., Sugiura, T., Ariyoshi, Y., Takahashi, T., and Ueda, R. (1993). Expression of the c-kit protein in human solid tumors and in corresponding fetal and adult normal tissues. Am. J. Pathol. 142, 339±346. Matsuguchi, T., Salgia, R., Hallek, M., Eder, M., Druker, B., Ernst, T. J., and Griffin, J. D. (1994). Shc phosphorylation in myeloid cells is regulated by granulocyte macrophage colonystimulating factor, interleukin-3, and steel factor and is constitutively increased by p210BCR=ABL . J. Biol. Chem. 269, 5016±5021. Matsuguchi, T., Inhorn, R. C., Carlesso, N., Xu, G., Druker, B., and Griffin, J. D. (1995). Tyrosine phosphorylation of p95Vav in myeloid cells is regulated by GM-CSF, IL-3 and Steel factor and is constitutively increased by p210BCR=ABL . EMBO J. 14, 257±265. Matsui, Y., Zsebo, K. M., and Hogan, B. L. (1990). Embryonic expression of a haematopoietic growth factor encoded by the S1 locus and the ligand for c-kit. Nature 347, 667±669. Mattei, S., Colombo, M. P., Melani, C., Silvani, A., Parmiani, G., and Herlyn, M. (1994). Expression of cytokine/growth factors and their receptors in human melanoma and melanocytes. Int. J. Cancer 56, 853±857. Melloti, P., and Calabretta, B. (1996). The transcription factors c-myb and GATA-2 act independently in the regulation of normal hematopoiesis. Proc. Natl Acad. Sci. USA 93, 5313±5318. Miller, B. A., Floros, J., Cheung, J. Y., Wojchowski, D. M., Bell, L., Begley, C. G., Elwood, N. J., Kreider, J., and Christian, C. (1994). Steel factor affects SCL expression during normal erythroid differentiation. Blood 84, 2971±2976. Miyazawa, K., Hendrie, P. C., Mantel, C., Wood, K., Ashman, L. K., and Broxmeyer, H. E. (1991). Comparative analysis of signaling pathways between mast cell growth factor (c-kit

ligand) and granulocyte-macrophage colony-stimulating factor in a human factor-dependent myeloid cell line involves phosphorylation of Raf-1, GTPase-activating protein and mitogenactivated protein kinase. Exp. Hematol. 19, 1110±1123. Miyazawa, K., Toyama, K., Gotoh, A., Hendrie, P. C., Mantel, C., and Broxmeyer, H. E. (1994). Ligand-dependent polyubiquitination of c-kit gene product: a possible mechanism of receptor down modulation in M07e cells. Blood 83, 137±145. Miyazawa, K., Williams, D. A., Gotoh, A., Nishimaki, J., Broxmeyer, H. E., and Toyama, K. (1995). Membrane-bound steel factor induces more persistent tyrosine kinase activation and longer life span of c-kit gene-encoded protein than its soluble form. Blood 85, 641±649. Montone, K. T., van Belle, P., Elenitsas, R., and Elder, D. E. (1997). Proto-oncogene c-kit expression in malignant melanoma: protein loss with tumor progression. Mod. Pathol. 10, 939±944. Morii, E., Hirota, S., Kim, H.-M., Mikoshiba, K., Nishimune, Y., Kitamura, Y., and Nomura, S. (1992). Spatial expression of genes encoding c-kit receptors and their ligands in mouse cerebellum as revealed by in situ hybridization. Dev. Brain Res. 65, 123±126. Morrison-Graham, K., and Takahashi, Y. (1993). Steel factor and c-Kit receptor: from mutants to a growth factor system. BioEssays 15, 77±83. Moskaluk, C. A., Tian, Q., Marshall, C. R., Rumpel, C. A., Franquemont, D. W., and Frierson Jr., H. F. (1999). Mutations of c-kit JM domain are found in a minority of human gastrointestinal stromal tumors. Oncogene 18, 1897± 1902. Motro, B., van der Kooy, D., Rossant, J., Reith, A., and Bernstein, A. (1991). Contiguous patterns of c-kit and steel expression: analysis of mutations at the W and S1 loci. Development 113, 1207±1221. Motro, B., Wojtowicz, J. M., Bernstein, A., and van der Kooy, D. (1996). Steel mutant mice are deficient in hippocampal learning but not long-term potentiation. Proc. Natl Acad. Sci. USA 93, 1808±1813. Muszynski, K. W., Ruscetti, F. W., Heidecker, G., Rapp, U., Troppmair, J., Gooya, J. M., and Keller, J. R. (1995). Raf-1 protein is required for growth factor-induced proliferation of hematopoietic cells. J. Exp. Med. 181, 2189±2199. Nagata, H., Worobec, A. S., Oh, C. K., Chowdhury, B. A., Tannenbaum, S., Suzuki, Y., and Metcalfe, D. D. (1995). Identification of a point mutation in the catalytic domain of the protooncogene c-kit in peripheral blood mononuclear cells of patients who have mastocytosis with an associated hematologic disorder. Proc. Natl Acad. Sci. USA 92, 10560±10564. Natali, P. G., Nicotra, M. R., Sures, I., Santoro, E., Bigotti, A., and Ullrich, A. (1992). Expression of c-kit receptor in normal and transformed human nonlymphoid tissues. Cancer Res. 52, 6139±6143. Natali, P. G., Berlingieri, M. T., Nicotra, M. R., Fusco, A., Santoro, E., Bigotti, A., and Vecchio, G. (1995). Transformation of thyroid epithelium is associated with loss of c-kit receptor. Cancer Res. 55, 1787±1791. Nilsson, G., Miettinen, U., Ishizaka, T., Ashman, L. K., Irani, A. M., and Schwartz, L. B. (1994). Interleukin-4 inhibits the expression of Kit and tryptase during stem cell factor-dependent development of human mast cells from fetal liver cells. Blood 84, 1519±1527. Nishikawa, S., Kusakabe, M., Yoshinaga, K., Ogawa, M., Hayashi, S.-I., Kunisada, T., Era, T., Sakakura, T., and Nishikawa, S.-I. (1991). In utero manipulation of coat color formation by a monoclonal anti-c-kit antibody: two distinct

SCF Receptor 1931 waves of c-kit-dependency during melanocyte development. EMBO J. 10, 2111±2118. Nocka, K., Tan, J. C., Chiu, E., Chu, T. Y., Ray, P., Traktman, P., and Besmer, P. (1990). Molecular bases of dominant negative and loss of function mutations at the murine c-kit/white spotting locus: W 37 , W V , W 41 and W. EMBO J. 9, 1805±1813. O'Farrell, A.-M., Ichihara, M., Mui, A.L.-F., and Miyajima, A. (1996). Signaling pathways activated in a unique mast cell line where interleukin-3 supports survival and stem cell factor is required for a proliferative response. Blood 87, 3655±3665. Ogawa, M., Matsuzaki, Y., Nishikawa, S., Hayashi, S.-I., Kunisada, T., Sudo, T., Kina, T., Nakuchi, H., and Nishikawa, S.-I. (1991). Expression and function of c-kit in hemopoietic progenitor cells. J. Exp. Med. 174, 63±71. Ohashi, A., Funasaka, Y., Ueda, M., and Ichihashi, M. (1996). c-KIT receptor expression in cutaneous malignant melanoma and benign melanocytic naevi. Melanoma Res. 6, 25±30. Okuda, K., Sanghera, J. S., Pelech, S. L., Kanakura, Y., Hallek, M., Griffin, J. D., and Druker, B. J. (1992). Granulocyte-macrophage colony-stimulating factor, interleukin-3, and Steel factor induce rapid tyrosine phosphorylation of p42 and p44 MAP kinase. Blood 79, 2880±2887. Okura, M., Maeda, H., Nishikawa, S., and Mizoguchi, M. (1995). Effects of monoclonal anti-c-kit antibody (ACK2) on melanocytes in newborn mice. J. Invest. Dermatol. 105, 322±328. Orr-Urtreger, A., Avivi, A., Zimmer, Y., Givol, D., Yarden, Y., and Lonai, P. (1990). Developmental expression of c-kit, a proto-oncogene encoded by the W locus. Development 109, 911±923. Park, G. H., Plummer III, H. K., and Krystal, G. W. (1998). Selective Sp1 binding is critical for maximal activity of the human c-kit promoter. Blood 92, 4138±4149. Paulson, R. F., Vesely, S., Siminovitch, K. A., and Bernstein, A. (1996). Signalling by the W/Kit receptor tyrosine kinase is negatively regulated in vivo by the protein tyrosine phosphatase Shp1. Nature Genet. 13, 309±315. Pearson, M. A., O'Farrell, A.-M., Dexter, T. M., Whetton, A. D., Owen-Lynch, P. J., and Heyworth, C. M. (1998). Investigation of the molecular mechanisms underlying growth factor synergy: the role of ERK 2 activation in synergy. Growth Factors 15, 293±306. Philo, J. S., Wen, J., Wypych, J., Schwartz, M. G., Mendiaz, E. A., and Langley, K. E. (1996). Human stem cell factor dimer forms a complex with two molecules of the extracellular domain of its receptor, kit. J. Biol. Chem. 271, 6895±6902. Piao, X., and Bernstein, A. (1996). A point mutation in the catalytic domain of c-kit induces growth factor independence, tumorigenicity, and differentiation of mast cells. Blood 87, 3117±3123. Piao, X., Paulson, R., van der Geer, P., Pawson, T., and Bernstein, A. (1996). Oncogenic mutation in the Kit receptor tyrosine kinase alters substrate specificity and induces degradation of the protein tyrosine phosphatase SHP-1. Proc. Natl Acad. Sci. USA 93, 14665±14669. Pignon, J.-M. (1997). C-kit mutations and mast cell disorders: a model of activating mutations of growth factor receptors. Hematol. Cell Ther. 39, 114±116. Pignon, J. M., Giraudier, S., Duquesnoy, P., Jouault, H., Imbert, M., Vainchenker, W., Vernant, J. P., and Tulliez, M. (1997). A new c-kit mutation in a case of aggressive mast cell disease. Br. J. Haematol. 96, 374±376. Plummer III, H., Catlett, J., Leftwich, J., Armstrong, B., Carlson, P., Huff, T., and Krystal, G. (1993). c-myc expression correlates with suppression of c-kit protooncogene expression in small cell lung cancer cell lines. Cancer Res. 53, 4337±4342.

Price, D. J., Rivnay, B., Fu, Y., Jiang, S., Avraham, S., and Avraham, H. (1997). Direct association of Csk homologous kinase (CHK) with the diphosphorylated site Tyr568=570 of the activated c-KIT. J. Biol. Chem. 272, 5915±5920. Price, E. R., Ding, H.-F., Badalian, T., Bhattachhary, S., Takemoto, C., Yao, T.-P., Hemesath, T. J., and Fisher, D. E. (1998). Lineage-specific signaling in melanocytes. J. Biol. Chem. 273, 17983±17986. Qiu, F., Ray, P., Brown, K., Barker, P. E., Jhanwar, S., Ruddle, F. H., and Besmer, P. (1988). Primary structure of c-kit: relationship with the CSF-1/PDGF receptor kinase family ± oncogenic activation of v-kit involves deletion of extracellular domain and C terminus. EMBO J. 7, 1003±1011. Ratajczak, M. Z., Luger, S. M., DeRiel, K., Abrahm, J., Calabretta, B., and Gewirtz, A. M. (1992). Role of the KIT protooncogene in normal and malignant human hematopoiesis. Proc. Natl Acad. Sci. USA 89, 1710±1714. Ratajczak, M. Z., Perrotti, D., Melotti, P., Powzaniuk, M., Calabretta, B., Onodera, K., Kregenow, D. A., Machalinski, B., and Gewirtz, A. M. (1998). Myb and Ets proteins are candidate regulators of c-kit expression in human hematopoietic cells. Blood 91, 1934±1946. Reddy, G. P., and Quesenberry, P. J. (1996). Stem cell factor enhances interleukin-3 dependent induction of 68-kD calmodulin-binding protein and thymidine kinase activity in NFS-60 cells. Blood 87, 3195±3202. Reith, A. D., Ellis, C., Lyman, S. D., Anderson, D. M., Williams, D. E., Bernstein, A., and Pawson, T. (1991). Signal transduction by normal isoforms and W mutant variants of the kit receptor tyrosine kinase. EMBO J. 10, 2451±2459. Ridings, J., Macardle, P. J., Byard, R. W., Skinner, J., and Zola, H. (1995). Cytokine receptor expression by solid tumours. Ther. Immunol. 2, 67±76. Rossi, P., Marziali, G., Albanesi, C., Charlesworth, A., Geremia, R., and Sorrentino, V. (1992). A novel c-kit transcript, potentially encoding a truncated receptor, originates within a kit gene intron in mouse spermatids. Dev. Biol. 152, 203±207. Rottapel, R., Reedijk, M., Williams, D. E., Lyman, S. D., Anderson, D. M., Pawson, T., and Bernstein, A. (1991). The Steel/W transduction pathway: kit autophosphorylation and its association with a unique subset of cytoplasmic signaling proteins is induced by the steel factor. Mol. Cell. Biol. 11, 3043±3051. Rukstalis, D. B. (1996). Molecular mechanisms of testicular carcinogenesis. World J. Urol. 14, 347±352. Russell, E. S. (1979). Hereditary anemias of the mouse: a review for geneticists. Adv. Genet. 20, 357±459. Rusten, L. S., Smeland, E. B., Jacobsen, F. W., Lien, E., Lesslauer, W., Loetscher, H., Dubois, C. M., and Jacobsen, S. E. (1994). Tumor necrosis factor-alpha inhibits stem cell factor-induced proliferation of human bone marrow progenitor cells in vitro. Role of p55 and p75 tumor necrosis factor receptors. J. Clin. Invest. 94, 165±172. Ryan, J. J., Huang, H., McReynolds, L. J., Shelburne, C., Hu-Li, J., Huff, T. F., and Paul, W. E. (1997). Stem cell factor activates STAT-5 DNA binding in IL-3-derived bone marrow mast cells. Exp. Hematol. 25, 357±362. Rygaard, K., Nakamura, T., and Spang-Thomsen, M. (1993). Expression of the proto-oncogenes c-met and c-kit and their ligands, hepatocyte growth factor/scatter factor and stem cell factor in SCLC cell lines and xenografts. Br. J. Cancer 67, 137±146. Saito, S., Enomoto, M., Sakakura, S., Ishii, Y., Sudo, T., and Ichijo, M. (1994). Localization of stem cell factor (SCF) and c-kit mRNA in human placental tissue and biological effects of

1932 Diana Linnekin and Jonathan R. Keller SCF on DNA synthesis in primary cultured cytotrophoblasts. Biochem. Biophys. Res. Commun. 205, 1762±1769. Sasaki, E., Okamura, H., Chikamune T., Kanai, Y., Watanabe, M., Naito, M., and Sakurai, M. (1993). Cloning and expression of the chicken c-kit proto-oncogene. Gene 128, 257±261. Sattler, M., and Salgia, R. (1998). Role of the adapter protein CRKL in signal transduction of normal hematopoietic and BCR/ABL-transformed cells. Leukemia 12, 637±644. Sattler, M., Salgia, R., Shrikhande, G., Verma, S., Pisick, E., Prasad, K. V. S., and Griffin, J. D. (1997). Steel factor induces tyrosine phosphorylation of CRKL and binding of CRKL to a complex containing c-Kit, phosphatidylinositol 3-kinase, and p120CBL . J. Biol. Chem. 272, 10248±10253. Schmidt, L., Duh, F.-M., Chen, F., Kishida, T., Glenn, G., Choyke, P., Scherer, S. W., Zhuang, Z., Lubensky, I., Dean, M., Allikmets, R., Chidambaram, A., Bergerheim, U. R., Feltis, J. T., Casadevall, C., Zamarron, A., Bernues, M., Richard, S., Lips, C. J. M., Walther, M. M., Tsui, L.-C., Geil, L., Orcutt, M. L., Stackhouse, T., Lipan, J., Slife, L., Brauch, H., Decker, J., Niehans, G., Hughson, M. D., Moch, H., Storkel, S., Lerman, M. I., Linehan, W. M., and Zbar, B. (1997). Germline and somatic mutations in the tyrosine kinase domain of the MET proto-oncogene in papillary renal carcinomas. Nature Genet. 16, 68±73. Schwarzenberger, P., Spence, S. E., Gooya, J. M., Michiel, D., Curiel, D. T., Ruscetti, F. W., and Keller, J. R. (1996). Targeted gene transfer to human hematopoietic progenitor cell lines through the c-kit receptor. Blood 87, 472±478. Sekido, Y., Obata, Y., Ueda, R., Hida, T., Suyama, M., Shimokata, K., Ariyoshi, Y., and Takahashi, T. (1991). Preferential expression of c-kit protooncogene transcripts in small cell lung cancer. Cancer Res. 51, 2416±2419. Serve, H., Yee, N. S., Stella, G., Sepp-Lorenzino, L., Tan, J. C., and Besmer, P. (1995). Differential roles of PI3-kinase and Kit tyrosine 821 in Kit receptor-mediated proliferation, survival and cell adhesion in mast cells. EMBO J. 14, 473±483. Sette, C., Bevilacqua, A., Bianchini, A., Mangia, F., Geremia, R., and Rossi, P. (1997). Parthenogenetic activation of mouse eggs by microinjection of a truncated c-kit tyrosine kinase present in spermatozoa. Development 124, 2267±2274. Sette, C., Bevilacqua, A., Geremia, R., and Rossi, P. (1998). Involvement of phospholipase C 1 in mouse egg activation induced by a truncated form of the c-kit tyrosine kinase present in spermatozoa. J. Cell Biol. 142, 1063±1074. Shepherd, P. R., Withers, D. J., and Siddle, K. (1998). Phosphoinositide 3-kinase: the key switch mechanism in insulin signalling. Biochem. J. 333, 471±490. Shimizu, Y., Ashman, L. K., Du, Z., and Schwartz, L. B. (1996). Internalization of kit together with stem cell factor on human fetal liver-derived mast cells. J. Immunol. 156, 3443±3449. Sillaber, C., Strobl, H., Bevec, D., Ashman, L. K., Butterfield, J. H., Lechner, K., Maurer, D., Bettelheim, P., and Valent, P. (1991). IL-4 regulates c-kit proto-oncogene product expression in human mast and myeloid progenitor cells. J. Immunol. 147, 4224±4228. Sperling, C., Schwartz, S., BuÈchner, T., Thiel, E., Ludwig, W.-D. (1997). Expression of the stem cell factor receptor c-kit (CD117) in acute leukemias. Haematologica 82, 617±621. Spritz, R. A. (1992). Lack of apparent hematologic abnormalities in human patients with c-kit (stem cell factor receptor) gene mutations. Blood 79, 2497±2499. Spritz, R. A. (1994). Molecular basis of human piebaldism. J. Invest. Derm. 103, 1375±1405.

Spritz, R. A. (1997). Piebaldism, Waardenburg syndrome, and related disorders of melanocyte development. Semin. Cutaneous Med. Surg. 16, 15±23. Sui, X., Krantz, S. B., You, M., and Zhao, Z. (1998). Synergistic activation of MAP kinase (ERK1/2) by erythropoietin and stem cell factor is essential for expanded erythropoiesis. Blood 92, 1142±1149. Takaoka, A., Toyota, M., Hinoda, Y., Itoh, F., Mita, H., Kakiuchi, H., Adachi, M., and Imai, K. (1997). Expression and identification of aberrant c-kit transcripts in human cancer cells. Cancer Lett. 115, 257±261. Tanaka, S., Yanagisawa, N., Tojo, H., Kim, Y. J., Tsujimura, T., Kitamura, Y., Sawasaki, T., and Tachi, C. (1997). Molecular cloning of cDNA encoding the c-kit receptor of Shiba goats and a novel alanine insertion specific to goats and sheep in the kinase insert region. Biochim. Biophys. Acta 1352, 151±155. Tang, B., Mano, H., Yi, T., and Ihle, J. N. (1994). Tec kinase associates with c-kit and is tyrosine phosphorylated and activated following stem cell factor binding. Mol. Cell. Biol. 14, 8432±8437. Taniguchi, S., Dai, C. H., Price, J. O., and Krantz, S. B. (1997). Interferon gamma downregulates stem cell factor and erythropoietin receptors but not insulin-like growth factor-I receptors in human erythroid colony-forming cells. Blood 90, 2244±2252. Tauchi, T., Boswell, H. S., Leibowitz, D., and Broxmeyer, H. E. (1994a). Coupling between p210bcr-abl and Shc and Grb2 adaptor proteins in hematopoietic cells permits growth factor receptor-independent link to Ras activation pathway. J. Exp. Med. 179, 167±175. Tauchi, T., Feng, G.-S., Marshall, M. S., Shen, R., Mantel, C., Pawson, T., and Broxmeyer, H. E. (1994b). The ubiquitously expressed Syp phosphatase interacts with c-kit and Grb2 in hematopoietic cells. J. Biol. Chem. 269, 25206±25211. Timokhina, I., Kissel, H., Stella, G., and Besmer, P. (1998). Kit signaling through PI 3-kinase and Src kinase pathways: an essential role for Rac1 and JNK activation in mast cell proliferation. EMBO J. 17, 6250±6262. Torihashi, S., Ward, S. M., Nishikawa, S.-I., Nishi, K., Kobayashi, S., and Sanders, K. M. (1995). C-kit-dependent development of interstitial cells and electrical activity in the murine gastrointestinal tract. Cell Tissue Res. 280, 97±111. Toyota, M., Hinoda, Y., Itoh, F., Takaoka, A., Imai, K., and Yachi, A. (1994). Complementary DNA cloning and characterization of truncated form of c-kit in human colon carcinoma cells. Cancer Res. 54, 273±275. Tsai, M., Chen, R.-H., Tam, S.-Y., Blenis, J., and Galli, S. J. (1993). Activation of MAP kinases, pp90rsk and pp70-S6 kinases in mouse mast cells by signaling through the c-kit receptor tyrosine kinase or FcRI: rapamycin inhibits activation of pp70-S6 kinase and proliferation in mouse mast cells. Eur. J. Immunol. 23, 3286±3291. Tsujimura, T. (1996). Role of c-kit receptor tyrosine kinase in the development, survival and neoplastic transformation of mast cells. Pathol. Int. 46, 933±938. Tsujimura, T., Hirota, S., Nomura, S., Niwa, Y., Yamazaki, M., Tono, T., Morii, E., Kim, H. M., Kondo, K., and Nishimune, Y. (1991). Characterization of Ws mutant allele of rats: a 12-base deletion in tyrosine kinase domain of c-kit gene. Blood 78, 1942± 1946. Tsujimura, T., Furitsu, T., Morimoto, M., Isozaki, K., Nomura, S., Matsuzawa, Y., Kitamura, Y., and Kanakura, Y. (1994). Ligand-independent activation of c-kit receptor tyrosine kinase in a murine mastocytoma cell line P-815 generated by a point mutation. Blood 83, 2619±2626.

SCF Receptor 1933 Tsujimura, T., Morii, E., Nozaki, M., Hashimoto, K., Moriyama, Y., Takebayashi, K., Kondo, T., Kanakura, Y., and Kitamura, Y. (1996a). Involvement of transcription factor encoded by the mi locus in the expression of c-kit receptor tyrosine kinase in cultured mast cells of mice. Blood 88, 1225±1233. Tsujimura, T., Morimoto, M., Hashimoto, K., Moriyama, Y., Kitayama, H., Matsuzawa, Y., Kitamura, Y., and Kanakura, Y. (1996b). Constitutive activation of c-kit in FMA3 murine mastocytoma cells caused by deletion of seven amino acids at the juxtamembrane domain. Blood 87, 273±283. Tsujimura, T., Kanakura, Y., and Kitamura, Y. (1997). Mechanisms of constitutive activation of c-kit receptor tyrosine kinase. Leukemia 11, 396±398. Tsujimura, T., Hashimoto, K., Kitayama, H., Ikeda, H., Sugahara, H., Matsumura, I., Kaisho, T., Terada, N., Kitamura, Y., and Kanakura, Y. (1999). Activating mutation in the catalytic domain of c-kit elicits hematopoietic transformation by receptor self-association not at the ligand-induced dimerization site. Blood 93, 1319±1329. Turner, A. M., Zsebo, K. M., Martin, F., Jacobsen, F. W., Bennett, L. G., and Broudy, V. C. (1992). Nonhematopoietic tumor cell lines express stem cell factor and display c-kit receptors. Blood 80, 374±381. Turner, A. M., Bennett, L. G., Lin, N. L., Wypych, J., Bartley, T. D., Hunt, R. W., Atkins, H. L., Langley, K. E., Parker, V., Martin, F., and Broudy, V. C. (1995). Identification and characterization of a soluble c-kit receptor produced by human hematopoietic cell lines. Blood 85, 2052± 2058. Vandenbark, G. R., deCastro, C. M., Taylor, H., Dew-Knight, S., and Kaufman, R. E. (1992). Cloning and structural analysis of the human c-kit gene. Oncogene 7, 1259±1266. Vliagoftis, H., Worobec, A. S., and Metcalfe, D. D. (1997). Updates on cells and cytokines: the protooncogene c-kit and c-kit ligand in human disease. J. Allergy Clin. Immunol. 100, 435±440. Vosseller, K., Stella, G., Yee, N. S., and Besmer, P. (1997). c-Kit receptor signaling through its phosphatidylinositide-30 -kinasebinding site and protein kinase C: role in mast cell enhancement of degranulation, adhesion, and membrane ruffling. Mol. Biol. Cell. 8, 909±922. Wehrle-Haller, B., and Weston, J. A. (1995). Soluble and cellbound forms of steel factor activity play distinct roles in melanocyte precursor dispersal and survival on the lateral neural crest migration pathway. Development 121, 731±742. Weiler, S. R., Mou, S., DeBerry, C. S., Keller, J. R., Ruscetti, F. W., Ferris, D. K., Longo, D. L., and Linnekin, D. (1996). JAK2 is associated with the c-kit proto-oncogene product and is phosphorylated in response to stem cell factor. Blood 87, 3688±3693. Weiler, S. R., Gooya, J. M., Ortiz, M., Tsai, S., Collins, S. J., and Keller, J. R. (1999). D3: a gene induced during myeloid cell differentiation of Linlo c-Kit‡ Sca-1‡ progenitor cells. Blood 93, 527±536. Welham, M. J., and Schrader, J. W. (1991). Modulation of c-kit mRNA and protein by hemopoietic growth factors. Mol. Cell. Biol. 11, 2901±2904. Wessely, O., Mellitzer, G., von Lindern, M., Levitzki, A., Gazit, A., Ischenko, I., Hayman, M. J., and Beug, H. (1997). Distinct roles of the receptor tyrosine kinases c-ErbB and c-Kit in regulating the balance between erythroid cell proliferation and differentiation. Cell Growth Differ. 8, 481±493. Wisniewski, D., Strife, A., Berman, E., and Clarkson, B. (1996). c-kit ligand stimulates tyrosine phosphorylation of a similar

pattern of phosphotyrosyl proteins in primary primitive normal hematopoietic progenitors that are constitutively phosphorylated in comparable primitive progenitors in chronic phase chronic myelogenous leukemia. Leukemia 10, 229±237. Worobec, A. S., Semere, T., Nagata, H., and Metcalfe, D. D. (1998). Clinical correlates of the presence of the Asp816Val c-kit mutation in the peripheral blood mononuclear cells of patients with mastocytosis. Cancer 83, 2120±2129. Wypych, J., Bennett, L. G., Schwartz, M. G., Clogston, C. L., Lu, H. S., Broudy, V. C., Bartley, T. D., Parker, V. P., and Langley, K. E. (1995). Soluble kit receptor in human serum. Blood 85, 66±73. Yajima, T., Kanda, T., Yoshiike, K., and Kitamura, Y. (1998). Retroviral vector targeting human cells via c-Kit-stem cell factor interaction. Hum. Gene Ther. 9, 779±787. Yamamoto, K., Tojo, A., Aoki, N., and Shibuya, M. (1993). Characterization of the promoter region of the human c-kit proto-oncogene. Jpn J. Cancer Res. 84, 1136±1144. Yamanashi, Y., and Baltimore, D. (1997). Identification of the Abl- and rasGAP-associated 62 kDa protein as a docking protein, Dok. Cell 88, 205±211. Yamataka, A., Yamataka, T., Lane, G. J., Kobayashi, H., Sueyoshi, N., and Miyano, T. (1998). Necrotizing enterocolitis and C-KIT. J. Pediatr. Surg. 33, 1682±1685. Yarden, Y., and Ullrich, A. (1988). Molecular analysis of signal transduction by growth factors. Biochemistry 27, 3113±3119. Yarden, Y., Kuang, W.-J., Yang-Feng, T., Coussens, L., Munemitsu, S., Dull, T. J., Chen, E., Schlessinger, J., Francke, U., and Ullrich, A. (1987). Human proto-oncogene c-kit: a new cell surface receptor tyrosine kinase for an unidentified ligand. EMBO J. 6, 3341±3351. Yasuda, H., Galli, S. J., and Geissler, E. N. (1993). Cloning and functional analysis of the mouse c-kit promoter. Biochem. Biophys. Res. Commun. 191, 893±901. Yee, N. S., Langen, H., and Besmer, P. (1993). Mechanism of kit ligand, phorbol ester, and calcium-induced down-regulation of c-kit receptors in mast cells. J. Biol. Chem. 268, 14289±14301. Yee, N. S., Hsiau, C.-W. M., Serve, H., Vosseller, K., and Besmer, P. (1994a). Mechanism of down-regulation of c-kit receptor. J. Biol. Chem. 269, 31991±31998. Yee, N. S., Paek, I., and Besmer, P. (1994b). Role of kit-ligand in proliferation and suppression of apoptosis in mast cells: basis for radiosensitivity of White Spotting and Steel mutant mice. J. Exp. Med. 179, 1777±1787. Yip-Schneider, M. T., Horie, M., and Broxmeyer, H. E. (1995). Transcriptional induction of pim-1 protein kinase gene expression by interferon and posttranscriptional effects on costimulation with steel factor. Blood 85, 3494±3502. Yoshida, H., Takakura, N., Kataoka, H., Kunisada, T., Okamura, H., and Nishikawa, S.-I. (1997). Stepwise requirement of c-kit tyrosine kinase in mouse ovarian follicle development. Dev. Biol. 184, 122±137. Zhang, Z., and Anthony, R. V. (1994). Porcine stem cell factor/ c-kit ligand: its molecular cloning and localization within the uterus. Biol. Reprod. 50, 95±102. Zhang, S.-C., and Fedoroff, S. (1997). Cellular localization of stem cell factor and c-kit receptor in the mouse nervous system. J. Neurosci. Res. 47, 1±15. Zhang, B. Y.-Y., Vik, T. A., Ryder, J. W., Srour, E. F., Jacks, T., Shannon, K., and Clapp, D. W. (1998). Nf1 regulates hematopoietic progenitor cell growth and Ras signaling in response to multiple cytokines. J. Exp. Med. 187, 1893±1902. Zsebo, K. M., Williams, D. A., Geissler, E. N., Broudy, V. C., Martin, F. H., Atkins, H. L., Hsu, R.-Y., Birkett, N. C., Okino, K. H., Murdock, D. C., Jacobsen, F. W.,

1934 Diana Linnekin and Jonathan R. Keller Langley, K. E., Smith, K. A., Takeishi, T., Cattanach, B. M., Galli, S. J., and Suggs, S. V. (1990). Stem cell factor is encoded at the S1 locus of the mouse and is the ligand for the c-kit tyrosine kinase receptor. Cell 63, 213±224.

been used to antagonize SCF-mediated proliferative responses. An inhibitor of the SCFR kinase activity, AG1296, is sold by Calbiochem.

LICENSED PRODUCTS

ACKNOWLEDGEMENTS

A variety of companies sell antisera specific for either human or murine SCFR. These products, the sources and some known uses are summarized in Table 4. A recombinant form of the SCFR extracellular domain is sold by R&D Systems. This product has

The authors would like to thank Dr Douglas Lowy, Dr RuJu Chian, Sherry Mou, and Bridget O'Laughlin for critical review of this manuscript.

Table 4 Commercially available antisera specific for the SCFR Species specificity

Monoclonal/ polyclonal

Epitope

Designation

Vendor

Use

Mouse

M

Binding site

ACK2

Gibco-BRL-LTI

FC, IB, N

Mouse

M

ECD

2B8

Pharmingen

FC

Mouse

M

ECD

ACK45

Pharmingen

FC, N

Mouse

P

C-tail

M-14

Santa Cruz

IP, IB

Mouse/human

P

C-tail

±

UBI

IP, IB

Mouse/human

P

C-tail

C-19

Santa Cruz

IP, IB

Human

M

Binding site

3D6

Boehringer-Mannheim

IP, IB, FC, N

Human

M

Binding site

K44.2

Sigma

IP, FC

Human

M

ECD

YB5.B8

Pharmingen

FC

Human

M

ECD

95C3

Coulter-Immunotech

FC

Human

M

ECD

17F11

Coulter-Immunotech

FC

Human

M

ECD

104D2D1

oulter-Immunotech

FC

Human

P

ECD

±

R&D

IB

Human

P

C-tail

C-14

Santa Cruz

IP, IB

Human

P

C-tail

±

Oncogene

IF

M, monoclonal; P, polyclonal; ECD, extracellular domain; C-tail, carboxyl tail; IB, immunoblotting; IP, immunoprecipitation; FC, flow cytometry; N, neutralizing.