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. 1985 Mar 1;100(3):748–753. doi: 10.1083/jcb.100.3.748

Effect upon mitogenic stimulation of calcium-dependent phosphorylation of cytoskeleton-associated 350,000- and 80,000-mol-wt polypeptides in quiescent 3Y1 cells

PMCID: PMC2113513  PMID: 3972893

Abstract

Rabbit antiserum raised against highest molecular weight microtubule- associated protein (MAP-1) of brain immunoprecipitated 350,000-, 300,000-, and 80,000-mol-wt phosphoproteins of rat embryo fibroblasts (3Y1-B). The 350,000-mol-wt protein was sensitive to heat as was brain MAP-1, but the 300,000- and 80,000-mol-wt proteins were not. These polypeptides were hardly phosphorylated in cells in the quiescent G0 phase but were rapidly phosphorylated after addition of serum, epidermal growth factor, phorbol ester, insulin, or transferrin in the presence of calcium ions. All these agents also induced incorporation of [3H]-thymidine into DNA. These polypeptides were detected in isolated microtubules and cold-resistant filaments by immunoblotting. Since the 350,000-mol-wt polypeptide was detected in the membrane, the cytoskeletons, and the nucleus, and has been suggested to function as a linker, its rapid phosphorylation might represent an early process in transduction of the signal of mitogenic stimulation to the nucleus.

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Selected References

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  1. Bennett V., Davis J. Erythrocyte ankyrin: immunoreactive analogues are associated with mitotic structures in cultured cells and with microtubules in brain. Proc Natl Acad Sci U S A. 1981 Dec;78(12):7550–7554. doi: 10.1073/pnas.78.12.7550. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Blat C., Harel L., Villaudy J., Golde A. Rous sarcoma virus-induced changes in the pattern of phosphorylation of non-histone nuclear proteins. Exp Cell Res. 1983 May;145(2):305–314. doi: 10.1016/0014-4827(83)90009-5. [DOI] [PubMed] [Google Scholar]
  3. Bloom G. S., Luca F. C., Vallee R. B. Widespread cellular distribution of MAP-1A (microtubule-associated protein 1A) in the mitotic spindle and on interphase microtubules. J Cell Biol. 1984 Jan;98(1):331–340. doi: 10.1083/jcb.98.1.331. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bockus B. J., Stiles C. D. Regulation of cytoskeletal architecture by platelet-derived growth factor, insulin and epidermal growth factor. Exp Cell Res. 1984 Jul;153(1):186–197. doi: 10.1016/0014-4827(84)90460-9. [DOI] [PubMed] [Google Scholar]
  5. Borisy G. G., Marcum J. M., Olmsted J. B., Murphy D. B., Johnson K. A. Purification of tubulin and associated high molecular weight proteins from porcine brain and characterization of microtubule assembly in vitro. Ann N Y Acad Sci. 1975 Jun 30;253:107–132. doi: 10.1111/j.1749-6632.1975.tb19196.x. [DOI] [PubMed] [Google Scholar]
  6. Cochet C., Gill G. N., Meisenhelder J., Cooper J. A., Hunter T. C-kinase phosphorylates the epidermal growth factor receptor and reduces its epidermal growth factor-stimulated tyrosine protein kinase activity. J Biol Chem. 1984 Feb 25;259(4):2553–2558. [PubMed] [Google Scholar]
  7. Connolly J. A., Kalnins V. I., Cleveland D. W., Kirschner M. W. Intracellular localization of the high molecular weight microtubule accessory protein by indirect immunofluorescence. J Cell Biol. 1978 Mar;76(3):781–786. doi: 10.1083/jcb.76.3.781. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Crossin K. L., Carney D. H. Evidence that microtubule depolymerization early in the cell cycle is sufficient to initiate DNA synthesis. Cell. 1981 Jan;23(1):61–71. doi: 10.1016/0092-8674(81)90270-1. [DOI] [PubMed] [Google Scholar]
  9. Friedkin M., Legg A., Rozengurt E. Antitubulin agents enhance the stimulation of DNA synthesis by polypeptide growth factors in 3T3 mouse fibroblasts. Proc Natl Acad Sci U S A. 1979 Aug;76(8):3909–3912. doi: 10.1073/pnas.76.8.3909. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Greene L. A., Liem R. K., Shelanski M. L. Regulation of a high molecular weight microtubule-associated protein in PC12 cells by nerve growth factor. J Cell Biol. 1983 Jan;96(1):76–83. doi: 10.1083/jcb.96.1.76. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Iwashita S., Fox C. F. Epidermal growth factor and potent phorbol tumor promoters induce epidermal growth factor receptor phosphorylation in a similar but distinctively different manner in human epidermoid carcinoma A431 cells. J Biol Chem. 1984 Feb 25;259(4):2559–2567. [PubMed] [Google Scholar]
  12. Kaibuchi K., Takai Y., Sawamura M., Hoshijima M., Fujikura T., Nishizuka Y. Synergistic functions of protein phosphorylation and calcium mobilization in platelet activation. J Biol Chem. 1983 Jun 10;258(11):6701–6704. [PubMed] [Google Scholar]
  13. Kessler S. W. Rapid isolation of antigens from cells with a staphylococcal protein A-antibody adsorbent: parameters of the interaction of antibody-antigen complexes with protein A. J Immunol. 1975 Dec;115(6):1617–1624. [PubMed] [Google Scholar]
  14. Klein I., Willingham M., Pastan I. A high molecular weight phosphoprotein in cultured fibroblasts that associates with polymerized tubulin. Exp Cell Res. 1978 Jun;114(1):229–238. doi: 10.1016/0014-4827(78)90056-3. [DOI] [PubMed] [Google Scholar]
  15. Kuznetsov S. A., Rodionov V. I., Gelfand V. I., Rosenblat V. A. Microtubule-associated protein MAP1 promotes microtubule assembly in vitro. FEBS Lett. 1981 Dec 7;135(2):241–244. doi: 10.1016/0014-5793(81)80791-0. [DOI] [PubMed] [Google Scholar]
  16. Nishida E., Kuwaki T., Sakai H. Phosphorylation of microtubule-associated proteins (MAPs) and pH of the medium control interaction between MAPs and actin filaments. J Biochem. 1981 Aug;90(2):575–578. doi: 10.1093/oxfordjournals.jbchem.a133510. [DOI] [PubMed] [Google Scholar]
  17. Nishizuka Y. The role of protein kinase C in cell surface signal transduction and tumour promotion. Nature. 1984 Apr 19;308(5961):693–698. doi: 10.1038/308693a0. [DOI] [PubMed] [Google Scholar]
  18. Okuda A., Kimura G. Effects of serum deprivation on the initiation of DNA synthesis in the second generation in rat 3Y1 cells. J Cell Physiol. 1982 Mar;110(3):267–270. doi: 10.1002/jcp.1041100308. [DOI] [PubMed] [Google Scholar]
  19. Otto A. M., Zumbé A., Gibson L., Kubler A. M., Jimenez de Asua L. Cytoskeleton-disrupting drugs enhance effect of growth factors and hormones on initiation of DNA synthesis. Proc Natl Acad Sci U S A. 1979 Dec;76(12):6435–6438. doi: 10.1073/pnas.76.12.6435. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Pytela R., Wiche G. High molecular weight polypeptides (270,000-340,000) from cultured cells are related to hog brain microtubule-associated proteins but copurify with intermediate filaments. Proc Natl Acad Sci U S A. 1980 Aug;77(8):4808–4812. doi: 10.1073/pnas.77.8.4808. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Rozengurt E., Rodriguez-Pena M., Smith K. A. Phorbol esters, phospholipase C, and growth factors rapidly stimulate the phosphorylation of a Mr 80,000 protein in intact quiescent 3T3 cells. Proc Natl Acad Sci U S A. 1983 Dec;80(23):7244–7248. doi: 10.1073/pnas.80.23.7244. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Sato C., Nishizawa K., Nakamura H., Komagoe Y., Shimada K., Ueda R., Suzuki S. Monoclonal antibody against microtubule associated protein-1 produces immunofluorescent spots in the nucleus and centrosome of cultured mammalian cells. Cell Struct Funct. 1983 Sep;8(3):245–254. doi: 10.1247/csf.8.245. [DOI] [PubMed] [Google Scholar]
  23. Sato C., Nishizawa K., Yamaguchi N. Co-localization of SV40 T antigen and p53 with immunological analogues of microtubule-associated protein-1 on the nuclear skeleton. Cell Struct Funct. 1984 Sep;9(3):305–309. doi: 10.1247/csf.9.305. [DOI] [PubMed] [Google Scholar]
  24. Sherline P., Schiavone K. Immunofluorescence localization of proteins of high molecular weight along intracellular microtubules. Science. 1977 Dec 9;198(4321):1038–1040. doi: 10.1126/science.337490. [DOI] [PubMed] [Google Scholar]
  25. Teng M. H., Bartholomew J. C., Bissell M. J. Synergism between anti-microtubule agents and growth stimulants in enhancement of cell cycle traverse. Nature. 1977 Aug 25;268(5622):739–741. doi: 10.1038/268739a0. [DOI] [PubMed] [Google Scholar]
  26. Vallee R. B., Davis S. E. Low molecular weight microtubule-associated proteins are light chains of microtubule-associated protein 1 (MAP 1). Proc Natl Acad Sci U S A. 1983 Mar;80(5):1342–1346. doi: 10.1073/pnas.80.5.1342. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Vasiliev J. M., Gelfand I. M., Guelstein V. I. Initiation of DNA synthesis in cell cultures by colcemid. Proc Natl Acad Sci U S A. 1971 May;68(5):977–979. doi: 10.1073/pnas.68.5.977. [DOI] [PMC free article] [PubMed] [Google Scholar]

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