CRK protein binds to two guanine nucleotide-releasing proteins for the Ras family and modulates nerve growth factor-induced activation of Ras in PC12 cells.

Matsuda, Minoru; Hashimoto, Yuko; Muroya, Kenkoh; Hasegawa, Hideki; Kurata, Takeshi; Tanaka, Shinya; Nakamura, Shun; Hattori, Satoshi

doi:10.1128/mcb.14.8.5495

Cited by 179 publications

(168 citation statements)

References 35 publications

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“…Crk binds to Sos, which acts on Ras Matsuda et al, 1994). However, it appears that Sos is a major e ector of another adaptor molecule, Grb2 (Buday and Downward, 1993;Lowenstein et al, 1992).…”

Section: Discussionmentioning

confidence: 99%

Identification of Tyr-762 in the platelet-derived growth factor α-receptor as the binding site for Crk proteins

et al. 1998

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Tyr-762 is an autophosphorylation site in the human platelet-derived growth factor (PDGF) a-receptor. In order to investigate whether phosphorylated Tyr-762 serves as a docking site for downstream signal transduction molecules, a nity puri®cation using an immobilized synthetic peptide containing phosphorylated Tyr-762 and its surrounding amino acid residues was performed. Proteins in HeLa cell lysate of molecular sizes 27, 38 and 40 kDa bound to the phosphorylated, but not to the unphosphorylated peptide. Analyses of partial amino acid sequences of the puri®ed proteins indicated that they were identical to CrkI, CrkII and CrkL respectively. The wild-type PDGF a-receptor, when expressed in porcine aortic endothelial cells, formed complexes with CrkII and CrkL upon ligand stimulation, which was speci®cally inhibited by a synthetic peptide containing phosphorylated Tyr-762. Replacement of Tyr-762 with a phenylalanine residue in the PDGF a-receptor abrogated ligand-induced binding of Crk proteins. Tyrosine phosphorylation of CrkII and CrkL increased by 1.8-and 1.3-fold, respectively, upon ligand stimulation of the wild-type areceptor. In contrast, the Y762F mutant PDGF areceptor failed to induce tyrosine phosphorylation of Crk proteins. CrkII and CrkL constitutively formed complex with the guanine nucleotide exchange factor C3G, in unstimulated as well as PDGF-stimulated cells. Moreover, the activated wild-type PDGF a-receptor but not the Y762F mutant receptor was found in a C3G immunoprecipitate, suggesting that a ternary complex between the activated PDGF a-receptor, Crk and C3G was formed. DNA synthesis stimulated by PDGF-BB as well as PDGF-induced MAP kinase activation was similar in cells expressing wild-type and mutant receptors. Interestingly, the activated PDGF b-receptor was found not to bind Crk proteins. Instead, Tyr-771 of the b-receptor, which is localized at an analogous position to Tyr-762 in the a-receptor, binds RasGAP. RasGAP is not bound to the a-receptor.

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Section: Discussionmentioning

confidence: 99%

Identification of Tyr-762 in the platelet-derived growth factor α-receptor as the binding site for Crk proteins

et al. 1998

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“…Expression of v-Crk results in an enhanced and sustained activation of Ras and MAP kinase after stimulation with EGF or NGF, implying that v-Crk can couple divergent tyrosine kinase pathways to Ras [44]. Overexpression of wildtype Crk-II, the cellular analogue of oncogenic v-Crk, in PC12 cells enhanced the NGF-induced activation of Ras, although the basal level of GTP-bound active Ras was not altered [29]. In contrast, mutants with a single amino acid substitution in either the SH2 or SH3 domains of the Crk protein inhibited the NGFinduced activation of Ras [34].…”

Section: Discussionmentioning

confidence: 99%

“…Despite the lack of a kinase domain these proteins are heavily phosphorylated in i o and in intact cells on tyrosine residues [27,28] and apparently have an important role in growth factor-stimulated signal transduction [28][29][30]. The widely expressed Crk-II protein contains an Nterminal SH2 domain followed by two SH3 domains [26,31].…”

Section: Introductionmentioning

confidence: 99%

Interaction in vitro of the product of the c-Crk-II proto-oncogene with the insulin-like growth factor I receptor

Koval

Blakesley

Roberts

et al. 1998

Biochemical Journal

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The Crk proto-oncogene product is an SH2 and SH3 domaincontaining adaptor protein. We have previously demonstrated that Crk-II becomes rapidly tyrosine-phosphorylated in response to stimulation with insulin-like growth factor I (IGF-I) and might be involved in the IGF-I receptor signalling pathway. To determine whether this involvement includes the direct interaction of Crk-II with the cytoplasmic region of the receptor, studies were performed in itro with glutathione S-transferase (GST) fusion proteins containing various domains of Crk-II. The kinase assay in itro showed that activated IGF-I receptors efficiently phosphorylated the GST-Crk-II fusion protein. This phosphorylation was dependent on the presence of the SH2 domain and Tyr-221 located in the spacer region between the two SH3 domains. Mutation of Tyr-221 not only prevented phosphorylation of GST-Crk in itro, but also significantly increased the ability of GST-Crk proteins to co-precipitate activated IGF-I receptors from total cell lysates. Additional binding experiments in itro showed that Crk-II might interact with the phosphorylated IGF-I receptor through its SH2 domain. To elucidate which

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“…It will therefore be important to compare the relative kinase activity of these pools of EGFR and to determine whether 70Z-Cbl has a greater or lesser e ect on their enhancement. Cbl's induced association with Crk proteins has also created interest because of the role of Crk in recruiting the nucleotide exchange factor C3G Matsuda et al, 1994) and therefore the activation of the Rap1A and Rap1B G-proteins (Gotoh et al, 1995). The putative role of Rap proteins as negative regulators of Ras (Hariharan et al, 1991;Cook et al, 1993) has obvious implications for providing a biochemical explanation for Sli-1's role as a negative regulator of receptor tyrosine kinases.…”

Section: Cbl Mediated-recruitment Of Crk To the Egfrmentioning

confidence: 99%

Tyrosine kinase activity of the EGF receptor is enhanced by the expression of oncogenic 70Z-Cbl

Thien

Langdon

1997

Oncogene

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The 120 kD product of the c-Cbl oncogene is a prominent substrate of protein tyrosine kinases that lacks a known catalytic activity but possesses an array of binding sites for cytoplasmic signalling proteins. An oncogenic form of Cbl was recently identi®ed in the 70Z/ 3 pre-B cell lymphoma which has a small deletion at the N-terminus of the Ring ®nger domain. This form of Cbl, termed 70Z-Cbl, exhibits an enhanced level of tyrosine phosphorylation compared with c-Cbl. Here we demonstrate that the expression of 70Z-Cbl induces a tenfold enhancement in the kinase activity of the EGF receptor in serum-starved and EGF-stimulated cells. In serumstarved cells this results in EGF receptor autophosphorylation and the recruitment of Grb2, Shc and Sos1 but does not induce a corresponding increase in MAP kinase activity. Furthermore the expression of 70Z-Cbl greatly enhances EGF-induced tyrosine phosphorylation of the protein tyrosine phosphatase SHP-2. We also show that the Cbl/EGF receptor complex is predominantly associated with CrkII and is distinct to the Grb2/ Shc/Sos1 complex that associates with the EGF receptor. These ®ndings therefore demonstrate a biochemical e ect of an oncogenic Cbl protein and support predictions from C. elegans that Cbl functions as regulator of receptor tyrosine kinases.

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CRK protein binds to two guanine nucleotide-releasing proteins for the Ras family and modulates nerve growth factor-induced activation of Ras in PC12 cells.

Cited by 179 publications

References 35 publications

Identification of Tyr-762 in the platelet-derived growth factor α-receptor as the binding site for Crk proteins

Identification of Tyr-762 in the platelet-derived growth factor α-receptor as the binding site for Crk proteins

Interaction in vitro of the product of the c-Crk-II proto-oncogene with the insulin-like growth factor I receptor

Tyrosine kinase activity of the EGF receptor is enhanced by the expression of oncogenic 70Z-Cbl

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