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. 1996 Oct 29;93(22):12177–12182. doi: 10.1073/pnas.93.22.12177

CrkII signals from epidermal growth factor receptor to Ras.

S Kizaka-Kondoh 1, M Matsuda 1, H Okayama 1
PMCID: PMC37963  PMID: 8901553

Abstract

A rat fibroblast mutant defective in oncogenic transformation and signaling from epidermal growth factor receptor to Ras has been isolated. The mutant contains dominant negative-type point mutations in the C-terminal SH3 domain of one crkII gene. Among the adapters tested, the mutant is complemented only by crkII cDNA. Expression of the mutated crkII in parent cells generates the phenotype indistinguishable from the mutant cell. Yet overexpression or reduced expression of Grb2 in the mutant before and after complementation with crkII have little effect on its phenotype. We conclude that adapter molecules are highly specific and that the oncogenic growth signal from epidermal growth factor receptor to Ras is predominantly mediated by CrkII in rat fibroblast.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Birge R. B., Fajardo J. E., Mayer B. J., Hanafusa H. Tyrosine-phosphorylated epidermal growth factor receptor and cellular p130 provide high affinity binding substrates to analyze Crk-phosphotyrosine-dependent interactions in vitro. J Biol Chem. 1992 May 25;267(15):10588–10595. [PubMed] [Google Scholar]
  2. Buday L., Downward J. Epidermal growth factor regulates p21ras through the formation of a complex of receptor, Grb2 adapter protein, and Sos nucleotide exchange factor. Cell. 1993 May 7;73(3):611–620. doi: 10.1016/0092-8674(93)90146-h. [DOI] [PubMed] [Google Scholar]
  3. Carpenter G., Cohen S. 125I-labeled human epidermal growth factor. Binding, internalization, and degradation in human fibroblasts. J Cell Biol. 1976 Oct;71(1):159–171. doi: 10.1083/jcb.71.1.159. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cerdá-Olmedo E., Hanawalt P. C., Guerola N. Mutagenesis of the replication point by nitrosoguanidine: map and pattern of replication of the Escherichia coli chromosome. J Mol Biol. 1968 May 14;33(3):705–719. doi: 10.1016/0022-2836(68)90315-x. [DOI] [PubMed] [Google Scholar]
  5. Chen C., Okayama H. High-efficiency transformation of mammalian cells by plasmid DNA. Mol Cell Biol. 1987 Aug;7(8):2745–2752. doi: 10.1128/mcb.7.8.2745. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Clark S. G., Stern M. J., Horvitz H. R. C. elegans cell-signalling gene sem-5 encodes a protein with SH2 and SH3 domains. Nature. 1992 Mar 26;356(6367):340–344. doi: 10.1038/356340a0. [DOI] [PubMed] [Google Scholar]
  7. Feller S. M., Ren R., Hanafusa H., Baltimore D. SH2 and SH3 domains as molecular adhesives: the interactions of Crk and Abl. Trends Biochem Sci. 1994 Nov;19(11):453–458. doi: 10.1016/0968-0004(94)90129-5. [DOI] [PubMed] [Google Scholar]
  8. Gale N. W., Kaplan S., Lowenstein E. J., Schlessinger J., Bar-Sagi D. Grb2 mediates the EGF-dependent activation of guanine nucleotide exchange on Ras. Nature. 1993 May 6;363(6424):88–92. doi: 10.1038/363088a0. [DOI] [PubMed] [Google Scholar]
  9. Kizaka-Kondoh S., Sato K., Tamura K., Nojima H., Okayama H. Raf-1 protein kinase is an integral component of the oncogenic signal cascade shared by epidermal growth factor and platelet-derived growth factor. Mol Cell Biol. 1992 Nov;12(11):5078–5086. doi: 10.1128/mcb.12.11.5078. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kume K., Jinno S., Miwatani H., Kizaka-Kondoh S., Terada Y., Nojima H., Okayama H. Oncogenic signal-induced ability to enter S phase in the absence of anchorage is the mechanism for the growth of transformed NRK cells in soft agar. New Biol. 1992 May;4(5):504–511. [PubMed] [Google Scholar]
  11. Lehmann J. M., Riethmüller G., Johnson J. P. Nck, a melanoma cDNA encoding a cytoplasmic protein consisting of the src homology units SH2 and SH3. Nucleic Acids Res. 1990 Feb 25;18(4):1048–1048. doi: 10.1093/nar/18.4.1048. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Li N., Batzer A., Daly R., Yajnik V., Skolnik E., Chardin P., Bar-Sagi D., Margolis B., Schlessinger J. Guanine-nucleotide-releasing factor hSos1 binds to Grb2 and links receptor tyrosine kinases to Ras signalling. Nature. 1993 May 6;363(6424):85–88. doi: 10.1038/363085a0. [DOI] [PubMed] [Google Scholar]
  13. Lowenstein E. J., Daly R. J., Batzer A. G., Li W., Margolis B., Lammers R., Ullrich A., Skolnik E. Y., Bar-Sagi D., Schlessinger J. The SH2 and SH3 domain-containing protein GRB2 links receptor tyrosine kinases to ras signaling. Cell. 1992 Aug 7;70(3):431–442. doi: 10.1016/0092-8674(92)90167-b. [DOI] [PubMed] [Google Scholar]
  14. Margolis B., Silvennoinen O., Comoglio F., Roonprapunt C., Skolnik E., Ullrich A., Schlessinger J. High-efficiency expression/cloning of epidermal growth factor-receptor-binding proteins with Src homology 2 domains. Proc Natl Acad Sci U S A. 1992 Oct 1;89(19):8894–8898. doi: 10.1073/pnas.89.19.8894. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Masuda A., Kizaka-Kondoh S., Miwatani H., Terada Y., Nojima H., Okayama H. Signal transduction cascade shared by epidermal growth factor and platelet-derived growth factor is a major pathway for oncogenic transformation in NRK cells. New Biol. 1992 May;4(5):489–503. [PubMed] [Google Scholar]
  16. Matsuda M., Hashimoto Y., Muroya K., Hasegawa H., Kurata T., Tanaka S., Nakamura S., Hattori S. 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. Mol Cell Biol. 1994 Aug;14(8):5495–5500. doi: 10.1128/mcb.14.8.5495. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Matsuda M., Tanaka S., Nagata S., Kojima A., Kurata T., Shibuya M. Two species of human CRK cDNA encode proteins with distinct biological activities. Mol Cell Biol. 1992 Aug;12(8):3482–3489. doi: 10.1128/mcb.12.8.3482. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Matuoka K., Shibata M., Yamakawa A., Takenawa T. Cloning of ASH, a ubiquitous protein composed of one Src homology region (SH) 2 and two SH3 domains, from human and rat cDNA libraries. Proc Natl Acad Sci U S A. 1992 Oct 1;89(19):9015–9019. doi: 10.1073/pnas.89.19.9015. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Mayer B. J., Hamaguchi M., Hanafusa H. A novel viral oncogene with structural similarity to phospholipase C. Nature. 1988 Mar 17;332(6161):272–275. doi: 10.1038/332272a0. [DOI] [PubMed] [Google Scholar]
  20. Meisenhelder J., Hunter T. The SH2/SH3 domain-containing protein Nck is recognized by certain anti-phospholipase C-gamma 1 monoclonal antibodies, and its phosphorylation on tyrosine is stimulated by platelet-derived growth factor and epidermal growth factor treatment. Mol Cell Biol. 1992 Dec;12(12):5843–5856. doi: 10.1128/mcb.12.12.5843. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Mizushima S., Nagata S. pEF-BOS, a powerful mammalian expression vector. Nucleic Acids Res. 1990 Sep 11;18(17):5322–5322. doi: 10.1093/nar/18.17.5322. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Muroya K., Hattori S., Nakamura S. Nerve growth factor induces rapid accumulation of the GTP-bound form of p21ras in rat pheochromocytoma PC12 cells. Oncogene. 1992 Feb;7(2):277–281. [PubMed] [Google Scholar]
  23. Ogawa S., Toyoshima H., Kozutsumi H., Hagiwara K., Sakai R., Tanaka T., Hirano N., Mano H., Yazaki Y., Hirai H. The C-terminal SH3 domain of the mouse c-Crk protein negatively regulates tyrosine-phosphorylation of Crk associated p130 in rat 3Y1 cells. Oncogene. 1994 Jun;9(6):1669–1678. [PubMed] [Google Scholar]
  24. Okayama H. Phage-mediated transduction of cDNA expression libraries into mammalian cells. Methods Enzymol. 1987;151:434–444. doi: 10.1016/s0076-6879(87)51035-7. [DOI] [PubMed] [Google Scholar]
  25. Olivier J. P., Raabe T., Henkemeyer M., Dickson B., Mbamalu G., Margolis B., Schlessinger J., Hafen E., Pawson T. A Drosophila SH2-SH3 adaptor protein implicated in coupling the sevenless tyrosine kinase to an activator of Ras guanine nucleotide exchange, Sos. Cell. 1993 Apr 9;73(1):179–191. doi: 10.1016/0092-8674(93)90170-u. [DOI] [PubMed] [Google Scholar]
  26. Pawson T. Protein modules and signalling networks. Nature. 1995 Feb 16;373(6515):573–580. doi: 10.1038/373573a0. [DOI] [PubMed] [Google Scholar]
  27. Reichman C. T., Mayer B. J., Keshav S., Hanafusa H. The product of the cellular crk gene consists primarily of SH2 and SH3 regions. Cell Growth Differ. 1992 Jul;3(7):451–460. [PubMed] [Google Scholar]
  28. Ren R., Ye Z. S., Baltimore D. Abl protein-tyrosine kinase selects the Crk adapter as a substrate using SH3-binding sites. Genes Dev. 1994 Apr 1;8(7):783–795. doi: 10.1101/gad.8.7.783. [DOI] [PubMed] [Google Scholar]
  29. Rozakis-Adcock M., Fernley R., Wade J., Pawson T., Bowtell D. The SH2 and SH3 domains of mammalian Grb2 couple the EGF receptor to the Ras activator mSos1. Nature. 1993 May 6;363(6424):83–85. doi: 10.1038/363083a0. [DOI] [PubMed] [Google Scholar]
  30. Schlessinger J., Bar-Sagi D. Activation of Ras and other signaling pathways by receptor tyrosine kinases. Cold Spring Harb Symp Quant Biol. 1994;59:173–179. doi: 10.1101/sqb.1994.059.01.021. [DOI] [PubMed] [Google Scholar]
  31. Simon M. A., Dodson G. S., Rubin G. M. An SH3-SH2-SH3 protein is required for p21Ras1 activation and binds to sevenless and Sos proteins in vitro. Cell. 1993 Apr 9;73(1):169–177. doi: 10.1016/0092-8674(93)90169-q. [DOI] [PubMed] [Google Scholar]
  32. Tanaka M., Gupta R., Mayer B. J. Differential inhibition of signaling pathways by dominant-negative SH2/SH3 adapter proteins. Mol Cell Biol. 1995 Dec;15(12):6829–6837. doi: 10.1128/mcb.15.12.6829. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Tanaka S., Morishita T., Hashimoto Y., Hattori S., Nakamura S., Shibuya M., Matuoka K., Takenawa T., Kurata T., Nagashima K. C3G, a guanine nucleotide-releasing protein expressed ubiquitously, binds to the Src homology 3 domains of CRK and GRB2/ASH proteins. Proc Natl Acad Sci U S A. 1994 Apr 12;91(8):3443–3447. doi: 10.1073/pnas.91.8.3443. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Teng K. K., Lander H., Fajardo J. E., Hanafusa H., Hempstead B. L., Birge R. B. v-Crk modulation of growth factor-induced PC12 cell differentiation involves the Src homology 2 domain of v-Crk and sustained activation of the Ras/mitogen-activated protein kinase pathway. J Biol Chem. 1995 Sep 1;270(35):20677–20685. doi: 10.1074/jbc.270.35.20677. [DOI] [PubMed] [Google Scholar]
  35. Tsuchie H., Chang C. H., Yoshida M., Vogt P. K. A newly isolated avian sarcoma virus, ASV-1, carries the crk oncogene. Oncogene. 1989 Nov;4(11):1281–1284. [PubMed] [Google Scholar]

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