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. 1984 Jan;4(1):30–37. doi: 10.1128/mcb.4.1.30

Diverse mitogenic agents induce the phosphorylation of two related 42,000-dalton proteins on tyrosine in quiescent chick cells.

J A Cooper, B M Sefton, T Hunter
PMCID: PMC368654  PMID: 6199661

Abstract

Incubation of quiescent chicken embryo cells with platelet-derived growth factor, epidermal growth factor, or serum was found to stimulate phosphorylation of two proteins of ca. 42,000 daltons on tyrosine. These proteins are structurally related to each other and to two proteins phosphorylated on tyrosine under similar conditions in mitogen-treated mouse fibroblasts. Three other very different mitogenic agents, the protease trypsin and the chemically unrelated tumor promoters 12-O-tetradecanoyl-phorbol-13-acetate and teleocidin, stimulated phosphorylation of the same proteins. In all cases, phosphotyrosine was detected in these phosphoproteins. Although additional changes in protein phosphorylation were evident, no other proteins were observed by two-dimensional gel electrophoresis which contained increased amounts of phosphotyrosine in mitogen-treated chicken embryo cells. One of these 42,000-dalton proteins was shown previously to be phosphorylated on tyrosine in chicken embryo cells transformed with various retroviruses whose transforming proteins possess tyrosine protein kinase activity. Phosphorylation of the 42,000-dalton proteins could be important in the regulation of cell division.

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Selected References

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

  1. Antoniades H. N., Stathakos D., Scher C. D. Isolation of a cationic polypeptide from human serum that stimulates proliferation of 3T3 cells. Proc Natl Acad Sci U S A. 1975 Jul;72(7):2635–2639. doi: 10.1073/pnas.72.7.2635. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Armelin H. A. Pituitary extracts and steroid hormones in the control of 3T3 cell growth. Proc Natl Acad Sci U S A. 1973 Sep;70(9):2702–2706. doi: 10.1073/pnas.70.9.2702. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Baker J. B., Low D. A., Simmer R. L., Cunningham D. D. Protease-nexin: a cellular component that links thrombin and plasminogen activator and mediates their binding to cells. Cell. 1980 Aug;21(1):37–45. doi: 10.1016/0092-8674(80)90112-9. [DOI] [PubMed] [Google Scholar]
  4. Bishop R., Martinez R., Nakamura K. D., Weber M. J. A tumor promoter stimulates phosphorylation on tyrosine. Biochem Biophys Res Commun. 1983 Sep 15;115(2):536–543. doi: 10.1016/s0006-291x(83)80178-8. [DOI] [PubMed] [Google Scholar]
  5. Bissell M. J., Hatié C., Calvin M. Is the product of the src gene a promoter? Proc Natl Acad Sci U S A. 1979 Jan;76(1):348–352. doi: 10.1073/pnas.76.1.348. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Blumberg P. M., Driedger P. E., Rossow P. W. Effect of a phorbol ester on a transformation-sensitive surface protein of chick fibroblasts. Nature. 1976 Dec 2;264(5585):446–447. doi: 10.1038/264446a0. [DOI] [PubMed] [Google Scholar]
  7. Bowen-Pope D. F., Ross R. Platelet-derived growth factor. II. Specific binding to cultured cells. J Biol Chem. 1982 May 10;257(9):5161–5171. [PubMed] [Google Scholar]
  8. Carney D. H., Cunningham D. D. Initiation of check cell division by trypsin action at the cell surface. Nature. 1977 Aug 18;268(5621):602–606. doi: 10.1038/268602a0. [DOI] [PubMed] [Google Scholar]
  9. Castagna M., Takai Y., Kaibuchi K., Sano K., Kikkawa U., Nishizuka Y. Direct activation of calcium-activated, phospholipid-dependent protein kinase by tumor-promoting phorbol esters. J Biol Chem. 1982 Jul 10;257(13):7847–7851. [PubMed] [Google Scholar]
  10. Chen L. B., Buchanan J. M. Mitogenic activity of blood components. I. Thrombin and prothrombin. Proc Natl Acad Sci U S A. 1975 Jan;72(1):131–135. doi: 10.1073/pnas.72.1.131. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Chen L. B., Buchanan J. M. Plasminogen-independent fibrinolysis by proteases produced by transformed chick embryo fibroblasts. Proc Natl Acad Sci U S A. 1975 Mar;72(3):1132–1136. doi: 10.1073/pnas.72.3.1132. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Cleveland D. W., Fischer S. G., Kirschner M. W., Laemmli U. K. Peptide mapping by limited proteolysis in sodium dodecyl sulfate and analysis by gel electrophoresis. J Biol Chem. 1977 Feb 10;252(3):1102–1106. [PubMed] [Google Scholar]
  13. Cohen S., Carpenter G., King L., Jr Epidermal growth factor-receptor-protein kinase interactions. Co-purification of receptor and epidermal growth factor-enhanced phosphorylation activity. J Biol Chem. 1980 May 25;255(10):4834–4842. [PubMed] [Google Scholar]
  14. Cooper J. A., Bowen-Pope D. F., Raines E., Ross R., Hunter T. Similar effects of platelet-derived growth factor and epidermal growth factor on the phosphorylation of tyrosine in cellular proteins. Cell. 1982 Nov;31(1):263–273. doi: 10.1016/0092-8674(82)90426-3. [DOI] [PubMed] [Google Scholar]
  15. Cooper J. A., Hunter T. Changes in protein phosphorylation in Rous sarcoma virus-transformed chicken embryo cells. Mol Cell Biol. 1981 Feb;1(2):165–178. doi: 10.1128/mcb.1.2.165. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Cooper J. A., Hunter T. Four different classes of retroviruses induce phosphorylation of tyrosines present in similar cellular proteins. Mol Cell Biol. 1981 May;1(5):394–407. doi: 10.1128/mcb.1.5.394. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Cooper J. A., Hunter T. Identification and characterization of cellular targets for tyrosine protein kinases. J Biol Chem. 1983 Jan 25;258(2):1108–1115. [PubMed] [Google Scholar]
  18. Cooper J. A., Reiss N. A., Schwartz R. J., Hunter T. Three glycolytic enzymes are phosphorylated at tyrosine in cells transformed by Rous sarcoma virus. Nature. 1983 Mar 17;302(5905):218–223. doi: 10.1038/302218a0. [DOI] [PubMed] [Google Scholar]
  19. Cooper J. A., Sefton B. M., Hunter T. Detection and quantification of phosphotyrosine in proteins. Methods Enzymol. 1983;99:387–402. doi: 10.1016/0076-6879(83)99075-4. [DOI] [PubMed] [Google Scholar]
  20. Cunningham D. D., Pardee A. B. Transport changes rapidly initiated by serum addition to "contact inhibited" 3T3 cells. Proc Natl Acad Sci U S A. 1969 Nov;64(3):1049–1056. doi: 10.1073/pnas.64.3.1049. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. De Asua L. J., Clingan D., Rudland P. S. Initiation of cell proliferation in cultured mouse fibroblasts by prostaglandin F2alpha. Proc Natl Acad Sci U S A. 1975 Jul;72(7):2724–2728. doi: 10.1073/pnas.72.7.2724. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Dicker P., Rozengurt E. Stimulation of DNA synthesis by tumour promoter and pure mitogenic factors. Nature. 1978 Dec 14;276(5689):723–726. doi: 10.1038/276723a0. [DOI] [PubMed] [Google Scholar]
  23. Dicker P., Rozengurt E. Tumour promoter induces proliferation of 3T6 cells in serum-free medium, as do polypeptide growth factors. Exp Cell Res. 1980 Dec;130(2):474–477. doi: 10.1016/0014-4827(80)90030-0. [DOI] [PubMed] [Google Scholar]
  24. Driedger P. E., Blumberg P. M. The effect of phorbol diesters on chicken embryo fibroblasts. Cancer Res. 1977 Sep;37(9):3257–3265. [PubMed] [Google Scholar]
  25. Ek B., Westermark B., Wasteson A., Heldin C. H. Stimulation of tyrosine-specific phosphorylation by platelet-derived growth factor. Nature. 1982 Feb 4;295(5848):419–420. doi: 10.1038/295419a0. [DOI] [PubMed] [Google Scholar]
  26. Erikson E., Erikson R. L. Identification of a cellular protein substrate phosphorylated by the avian sarcoma virus-transforming gene product. Cell. 1980 Oct;21(3):829–836. doi: 10.1016/0092-8674(80)90446-8. [DOI] [PubMed] [Google Scholar]
  27. Garrels J. I. Two dimensional gel electrophoresis and computer analysis of proteins synthesized by clonal cell lines. J Biol Chem. 1979 Aug 25;254(16):7961–7977. [PubMed] [Google Scholar]
  28. Gill G. N., Lazar C. S. Increased phosphotyrosine content and inhibition of proliferation in EGF-treated A431 cells. Nature. 1981 Sep 24;293(5830):305–307. doi: 10.1038/293305a0. [DOI] [PubMed] [Google Scholar]
  29. Glenn K. C., Cunningham D. D. Thrombin-stimulated cell division involves proteolysis of its cell surface receptor. Nature. 1979 Apr 19;278(5706):711–714. doi: 10.1038/278711a0. [DOI] [PubMed] [Google Scholar]
  30. Goldberg A. R., Delclos K. B., Blumberg P. M. Phorbol ester action is independent of viral and cellular src kinase levels. Science. 1980 Apr 11;208(4440):191–193. doi: 10.1126/science.6244621. [DOI] [PubMed] [Google Scholar]
  31. Gospodarowicz D. Localisation of a fibroblast growth factor and its effect alone and with hydrocortisone on 3T3 cell growth. Nature. 1974 May 10;249(453):123–127. doi: 10.1038/249123a0. [DOI] [PubMed] [Google Scholar]
  32. Hollenberg M. D., Cuatrecasas P. Epidermal growth factor: receptors in human fibroblasts and modulation of action by cholera toxin. Proc Natl Acad Sci U S A. 1973 Oct;70(10):2964–2968. doi: 10.1073/pnas.70.10.2964. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Hunter T., Cooper J. A. Epidermal growth factor induces rapid tyrosine phosphorylation of proteins in A431 human tumor cells. Cell. 1981 Jun;24(3):741–752. doi: 10.1016/0092-8674(81)90100-8. [DOI] [PubMed] [Google Scholar]
  34. Imai Y., Kaneko Y., Matsuzaki F., Endo Y., Oda T. Teleocidin B inhibits binding of epidermal growth factor to cellular receptors probably by the same mechanism as phorbol esters. Biochem Biophys Res Commun. 1980 Dec 16;97(3):926–931. doi: 10.1016/0006-291x(80)91465-5. [DOI] [PubMed] [Google Scholar]
  35. Kohler N., Lipton A. Platelets as a source of fibroblast growth-promoting activity. Exp Cell Res. 1974 Aug;87(2):297–301. doi: 10.1016/0014-4827(74)90484-4. [DOI] [PubMed] [Google Scholar]
  36. Laszlo A., Radke K., Chin S., Bissell M. J. Tumor promoters alter gene expression and protein phosphorylation in avian cells in culture. Proc Natl Acad Sci U S A. 1981 Oct;78(10):6241–6245. doi: 10.1073/pnas.78.10.6241. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Martinez R., Nakamura K. D., Weber M. J. Identification of phosphotyrosine-containing proteins in untransformed and Rous sarcoma virus-transformed chicken embryo fibroblasts. Mol Cell Biol. 1982 Jun;2(6):653–665. doi: 10.1128/mcb.2.6.653. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Nakamura K. D., Martinez R., Weber M. J. Tyrosine phosphorylation of specific proteins after mitogen stimulation of chicken embryo fibroblasts. Mol Cell Biol. 1983 Mar;3(3):380–390. doi: 10.1128/mcb.3.3.380. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Niedel J. E., Kuhn L. J., Vandenbark G. R. Phorbol diester receptor copurifies with protein kinase C. Proc Natl Acad Sci U S A. 1983 Jan;80(1):36–40. doi: 10.1073/pnas.80.1.36. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Nishimura J., Huang J. S., Deuel T. F. Platelet-derived growth factor stimulates tyrosine-specific protein kinase activity in Swiss mouse 3T3 cell membranes. Proc Natl Acad Sci U S A. 1982 Jul;79(14):4303–4307. doi: 10.1073/pnas.79.14.4303. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Pruss R. M., Herschman H. R. Variants of 3T3 cells lacking mitogenic response to epidermal growth factor. Proc Natl Acad Sci U S A. 1977 Sep;74(9):3918–3921. doi: 10.1073/pnas.74.9.3918. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Radke K., Gilmore T., Martin G. S. Transformation by Rous sarcoma virus: a cellular substrate for transformation-specific protein phosphorylation contains phosphotyrosine. Cell. 1980 Oct;21(3):821–828. doi: 10.1016/0092-8674(80)90445-6. [DOI] [PubMed] [Google Scholar]
  43. Raines E. W., Ross R. Platelet-derived growth factor. I. High yield purification and evidence for multiple forms. J Biol Chem. 1982 May 10;257(9):5154–5160. [PubMed] [Google Scholar]
  44. Reynolds F. H., Jr, Todaro G. J., Fryling C., Stephenson J. R. Human transforming growth factors induce tyrosine phosphorylation of EGF receptors. Nature. 1981 Jul 16;292(5820):259–262. doi: 10.1038/292259a0. [DOI] [PubMed] [Google Scholar]
  45. Rifkin D. B., Crowe R. M., Pollack R. Tumor promoters induce changes in the chick embryo fibroblast cytoskeleton. Cell. 1979 Oct;18(2):361–368. doi: 10.1016/0092-8674(79)90055-2. [DOI] [PubMed] [Google Scholar]
  46. Ross R., Glomset J., Kariya B., Harker L. A platelet-dependent serum factor that stimulates the proliferation of arterial smooth muscle cells in vitro. Proc Natl Acad Sci U S A. 1974 Apr;71(4):1207–1210. doi: 10.1073/pnas.71.4.1207. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Rozengurt E., Heppel L. A. Serum rapidly stimulates ouabain-sensitive 86-RB+ influx in quiescent 3T3 cells. Proc Natl Acad Sci U S A. 1975 Nov;72(11):4492–4495. doi: 10.1073/pnas.72.11.4492. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Rubin H. Overgrowth stimulating factor released from Rous sarcoma cells. Science. 1970 Feb 27;167(3922):1271–1272. doi: 10.1126/science.167.3922.1271. [DOI] [PubMed] [Google Scholar]
  49. Savage C. R., Jr, Cohen S. Epidermal growth factor and a new derivative. Rapid isolation procedures and biological and chemical characterization. J Biol Chem. 1972 Dec 10;247(23):7609–7611. [PubMed] [Google Scholar]
  50. Sefton B. M., Rubin H. Release from density dependent growth inhibition by proteolytic enzymes. Nature. 1970 Aug 22;227(5260):843–845. doi: 10.1038/227843a0. [DOI] [PubMed] [Google Scholar]
  51. Sefton B. M., Rubin H. Stimulation of glucose transport in cultures of density-inhibited chick embryo cells. Proc Natl Acad Sci U S A. 1971 Dec;68(12):3154–3157. doi: 10.1073/pnas.68.12.3154. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Sivak A. Induction of cell division: role of cell membrane sites. J Cell Physiol. 1972 Oct;80(2):167–173. doi: 10.1002/jcp.1040800203. [DOI] [PubMed] [Google Scholar]
  53. TODARO G. J., GREEN H., GOLDBERG B. D. TRANSFORMATION OF PROPERTIES OF AN ESTABLISHED CELL LINE BY SV40 AND POLYOMA VIRUS. Proc Natl Acad Sci U S A. 1964 Jan;51:66–73. doi: 10.1073/pnas.51.1.66. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Temin H. M. Studies on carcinogenesis by avian sarcoma viruses. VI. Differential multiplication of uninfected and of converted cells in response to insulin. J Cell Physiol. 1967 Jun;69(3):377–384. doi: 10.1002/jcp.1040690314. [DOI] [PubMed] [Google Scholar]
  55. Teng N. N., Bo Chen L. The role of surface proteins in cell proliferation as studied with thrombin and other proteases. Proc Natl Acad Sci U S A. 1975 Feb;72(2):413–417. doi: 10.1073/pnas.72.2.413. [DOI] [PMC free article] [PubMed] [Google Scholar]
  56. Thomas G., Siegmann M., Gordon J. Multiple phosphorylation of ribosomal protein S6 during transition of quiescent 3T3 cells into early G1, and cellular compartmentalization of the phosphate donor. Proc Natl Acad Sci U S A. 1979 Aug;76(8):3952–3956. doi: 10.1073/pnas.76.8.3952. [DOI] [PMC free article] [PubMed] [Google Scholar]
  57. 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]
  58. Unkeless J. C., Tobia A., Ossowski L., Quigley J. P., Rifkin D. B., Reich E. An enzymatic function associated with transformation of fibroblasts by oncogenic viruses. I. Chick embryo fibroblast cultures transformed by avian RNA tumor viruses. J Exp Med. 1973 Jan 1;137(1):85–111. doi: 10.1084/jem.137.1.85. [DOI] [PMC free article] [PubMed] [Google Scholar]
  59. Ushiro H., Cohen S. Identification of phosphotyrosine as a product of epidermal growth factor-activated protein kinase in A-431 cell membranes. J Biol Chem. 1980 Sep 25;255(18):8363–8365. [PubMed] [Google Scholar]
  60. Weber M. J., Rubin H. Uridine transport and RNA synthesis in growing and in density-inhibited animal cells. J Cell Physiol. 1971 Apr;77(2):157–168. doi: 10.1002/jcp.1040770205. [DOI] [PubMed] [Google Scholar]
  61. Weinstein I. B. Protein kinase, phospholipid and control of growth. Nature. 1983 Apr 28;302(5911):750–750. doi: 10.1038/302750a0. [DOI] [PubMed] [Google Scholar]
  62. Wigler M., Weinstein I. B. Tumour promotor induces plasminogen activator. Nature. 1976 Jan 22;259(5540):232–233. doi: 10.1038/259232a0. [DOI] [PubMed] [Google Scholar]

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