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. 1993 Apr 1;121(1):113–119. doi: 10.1083/jcb.121.1.113

Interference with endogenous ras function inhibits cellular responses to wounding

PMCID: PMC2119762  PMID: 8458864

Abstract

Wounding of tissue induces cellular responses that ultimately result in wound repair. Studies in tissue culture model systems indicate that these responses include induction of AP-1 regulated genes, cell migration and mitogenesis which are also characteristic of cellular responses to growth factors. Investigations have identified cellular ras proteins as critical components of growth factor-stimulated signal transduction pathways, however their role in the wounding response is less clear. Investigation of the potential involvement of c-Ras in this process utilized quiescent living bovine corneal endothelium cells (BCE) which were microinjected with ras dominant interfering mutant protein (N17) and subsequently stimulated by mechanical wounding. Analysis of these cells demonstrated that microinjection of dominant- interfering ras protein, but not control proteins, inhibited the wounding response as evidenced by diminished Fos expression, lack of cell migration and a block in DNA synthesis.

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Selected References

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  1. Bailleul B., Surani M. A., White S., Barton S. C., Brown K., Blessing M., Jorcano J., Balmain A. Skin hyperkeratosis and papilloma formation in transgenic mice expressing a ras oncogene from a suprabasal keratin promoter. Cell. 1990 Aug 24;62(4):697–708. doi: 10.1016/0092-8674(90)90115-u. [DOI] [PubMed] [Google Scholar]
  2. Bar-Sagi D., Feramisco J. R. Induction of membrane ruffling and fluid-phase pinocytosis in quiescent fibroblasts by ras proteins. Science. 1986 Sep 5;233(4768):1061–1068. doi: 10.1126/science.3090687. [DOI] [PubMed] [Google Scholar]
  3. Cai H., Szeberényi J., Cooper G. M. Effect of a dominant inhibitory Ha-ras mutation on mitogenic signal transduction in NIH 3T3 cells. Mol Cell Biol. 1990 Oct;10(10):5314–5323. doi: 10.1128/mcb.10.10.5314. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chiao P. J., Kannan P., Yim S. O., Krizman D. B., Wu T. A., Gallick G. E., Tainsky M. A. Susceptibility to ras oncogene transformation is coregulated with signal transduction through growth factor receptors. Oncogene. 1991 May;6(5):713–720. [PubMed] [Google Scholar]
  5. Deng G. R., Liu X. H., Wang J. R. Correlation of mutations of oncogene C-Ha-ras at codon 12 with metastasis and survival of gastric cancer patients. Oncogene Res. 1991;6(1):33–38. [PubMed] [Google Scholar]
  6. Dulbecco R., Stoker M. G. Conditions determining initiation of DNA synthesis in 3T3 cells. Proc Natl Acad Sci U S A. 1970 May;66(1):204–210. doi: 10.1073/pnas.66.1.204. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Farnsworth C. L., Feig L. A. Dominant inhibitory mutations in the Mg(2+)-binding site of RasH prevent its activation by GTP. Mol Cell Biol. 1991 Oct;11(10):4822–4829. doi: 10.1128/mcb.11.10.4822. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Feig L. A., Cooper G. M. Inhibition of NIH 3T3 cell proliferation by a mutant ras protein with preferential affinity for GDP. Mol Cell Biol. 1988 Aug;8(8):3235–3243. doi: 10.1128/mcb.8.8.3235. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Feldman S. T., Gately D., Schönthal A., Feramisco J. R. Fos expression and growth regulation in bovine corneal endothelial cells. Invest Ophthalmol Vis Sci. 1992 Nov;33(12):3307–3314. [PubMed] [Google Scholar]
  10. Gross M., Sweet R. W., Sathe G., Yokoyama S., Fasano O., Goldfarb M., Wigler M., Rosenberg M. Purification and characterization of human H-ras proteins expressed in Escherichia coli. Mol Cell Biol. 1985 May;5(5):1015–1024. doi: 10.1128/mcb.5.5.1015. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Guan J. L., Shalloway D. Regulation of focal adhesion-associated protein tyrosine kinase by both cellular adhesion and oncogenic transformation. Nature. 1992 Aug 20;358(6388):690–692. doi: 10.1038/358690a0. [DOI] [PubMed] [Google Scholar]
  12. Hunter T. Cooperation between oncogenes. Cell. 1991 Jan 25;64(2):249–270. doi: 10.1016/0092-8674(91)90637-e. [DOI] [PubMed] [Google Scholar]
  13. Hynes R. O. Integrins: versatility, modulation, and signaling in cell adhesion. Cell. 1992 Apr 3;69(1):11–25. doi: 10.1016/0092-8674(92)90115-s. [DOI] [PubMed] [Google Scholar]
  14. Ingber D. Integrins as mechanochemical transducers. Curr Opin Cell Biol. 1991 Oct;3(5):841–848. doi: 10.1016/0955-0674(91)90058-7. [DOI] [PubMed] [Google Scholar]
  15. Kyprianou N., Isaacs J. T. Relationship between metastatic ability and H-ras oncogene expression in rat mammary cancer cells transfected with the v-H-ras oncogene. Cancer Res. 1990 Mar 1;50(5):1449–1454. [PubMed] [Google Scholar]
  16. LaMorte V. J., Goldsmith P. K., Spiegel A. M., Meinkoth J. L., Feramisco J. R. Inhibition of DNA synthesis in living cells by microinjection of Gi2 antibodies. J Biol Chem. 1992 Jan 15;267(2):691–694. [PubMed] [Google Scholar]
  17. McKenna W. G., Weiss M. C., Bakanauskas V. J., Sandler H., Kelsten M. L., Biaglow J., Tuttle S. W., Endlich B., Ling C. C., Muschel R. J. The role of the H-ras oncogene in radiation resistance and metastasis. Int J Radiat Oncol Biol Phys. 1990 Apr;18(4):849–859. doi: 10.1016/0360-3016(90)90407-b. [DOI] [PubMed] [Google Scholar]
  18. McNeil P. L., Muthukrishnan L., Warder E., D'Amore P. A. Growth factors are released by mechanically wounded endothelial cells. J Cell Biol. 1989 Aug;109(2):811–822. doi: 10.1083/jcb.109.2.811. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Plantefaber L. C., Hynes R. O. Changes in integrin receptors on oncogenically transformed cells. Cell. 1989 Jan 27;56(2):281–290. doi: 10.1016/0092-8674(89)90902-1. [DOI] [PubMed] [Google Scholar]
  20. Ridley A. J., Paterson H. F., Johnston C. L., Diekmann D., Hall A. The small GTP-binding protein rac regulates growth factor-induced membrane ruffling. Cell. 1992 Aug 7;70(3):401–410. doi: 10.1016/0092-8674(92)90164-8. [DOI] [PubMed] [Google Scholar]
  21. Ruoslahti E., Yamaguchi Y. Proteoglycans as modulators of growth factor activities. Cell. 1991 Mar 8;64(5):867–869. doi: 10.1016/0092-8674(91)90308-l. [DOI] [PubMed] [Google Scholar]
  22. Sato Y., Rifkin D. B. Autocrine activities of basic fibroblast growth factor: regulation of endothelial cell movement, plasminogen activator synthesis, and DNA synthesis. J Cell Biol. 1988 Sep;107(3):1199–1205. doi: 10.1083/jcb.107.3.1199. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Schuh A. C., Keating S. J., Monteclaro F. S., Vogt P. K., Breitman M. L. Obligatory wounding requirement for tumorigenesis in v-jun transgenic mice. Nature. 1990 Aug 23;346(6286):756–760. doi: 10.1038/346756a0. [DOI] [PubMed] [Google Scholar]
  24. Settleman J., Narasimhan V., Foster L. C., Weinberg R. A. Molecular cloning of cDNAs encoding the GAP-associated protein p190: implications for a signaling pathway from ras to the nucleus. Cell. 1992 May 1;69(3):539–549. doi: 10.1016/0092-8674(92)90454-k. [DOI] [PubMed] [Google Scholar]
  25. Sheng M., Greenberg M. E. The regulation and function of c-fos and other immediate early genes in the nervous system. Neuron. 1990 Apr;4(4):477–485. doi: 10.1016/0896-6273(90)90106-p. [DOI] [PubMed] [Google Scholar]
  26. Shou C., Farnsworth C. L., Neel B. G., Feig L. A. Molecular cloning of cDNAs encoding a guanine-nucleotide-releasing factor for Ras p21. Nature. 1992 Jul 23;358(6384):351–354. doi: 10.1038/358351a0. [DOI] [PubMed] [Google Scholar]
  27. Stacey D. W., Roudebush M., Day R., Mosser S. D., Gibbs J. B., Feig L. A. Dominant inhibitory Ras mutants demonstrate the requirement for Ras activity in the action of tyrosine kinase oncogenes. Oncogene. 1991 Dec;6(12):2297–2304. [PubMed] [Google Scholar]
  28. Thurston G., Jaggi B., Palcic B. Measurement of cell motility and morphology with an automated microscope system. Cytometry. 1988 Sep;9(5):411–417. doi: 10.1002/cyto.990090502. [DOI] [PubMed] [Google Scholar]
  29. Todaro G. J., Lazar G. K., Green H. The initiation of cell division in a contact-inhibited mammalian cell line. J Cell Physiol. 1965 Dec;66(3):325–333. doi: 10.1002/jcp.1030660310. [DOI] [PubMed] [Google Scholar]
  30. Trahey M., Milley R. J., Cole G. E., Innis M., Paterson H., Marshall C. J., Hall A., McCormick F. Biochemical and biological properties of the human N-ras p21 protein. Mol Cell Biol. 1987 Jan;7(1):541–544. doi: 10.1128/mcb.7.1.541. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Verrier B., Müller D., Bravo R., Müller R. Wounding a fibroblast monolayer results in the rapid induction of the c-fos proto-oncogene. EMBO J. 1986 May;5(5):913–917. doi: 10.1002/j.1460-2075.1986.tb04303.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Werb Z., Tremble P. M., Behrendtsen O., Crowley E., Damsky C. H. Signal transduction through the fibronectin receptor induces collagenase and stromelysin gene expression. J Cell Biol. 1989 Aug;109(2):877–889. doi: 10.1083/jcb.109.2.877. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Yayon A., Klagsbrun M. Autocrine transformation by chimeric signal peptide-basic fibroblast growth factor: reversal by suramin. Proc Natl Acad Sci U S A. 1990 Jul;87(14):5346–5350. doi: 10.1073/pnas.87.14.5346. [DOI] [PMC free article] [PubMed] [Google Scholar]

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