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. 1986 Jun;83(12):4190–4193. doi: 10.1073/pnas.83.12.4190

Phosphorylation of tyrosine residues of calmodulin in Rous sarcoma virus-transformed cells.

Y Fukami, T Nakamura, A Nakayama, T Kanehisa
PMCID: PMC323697  PMID: 2424020

Abstract

Calmodulin, a wide-spread eukaryotic Ca2+-binding protein, was phosphorylated at its tyrosine residues in Rous sarcoma virus (RSV)-transformed chicken and rat cells but not in normal chicken embryo fibroblasts. In contrast, serine and threonine phosphorylation of calmodulin was found to occur in both normal and virus-transformed cells. In an in vitro system containing purified src kinase from RSV-transformed cells, tyrosine phosphorylation of calmodulin by the src kinase was inhibited by Ca2+. Furthermore, the tyrosine-phosphorylated calmodulin showed slower mobility than that of nonphosphorylated calmodulin in NaDodSO4/polyacrylamide gel electrophoresis when Ca2+ was present. These results suggest that the structure of calmodulin Ca2+ complex may be altered by tyrosine phosphorylation. It is thus inferred that Ca2+ may regulate the level of tyrosine phosphorylation of calmodulin in RSV-transformed cells, and phosphorylation in turn may attenuate the function of this protein in vivo.

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Selected References

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

  1. Burgess W. H., Jemiolo D. K., Kretsinger R. H. Interaction of calcium and calmodulin in the presence of sodium dodecyl sulfate. Biochim Biophys Acta. 1980 Jun 26;623(2):257–270. doi: 10.1016/0005-2795(80)90254-8. [DOI] [PubMed] [Google Scholar]
  2. Collett M. S., Erikson R. L. Protein kinase activity associated with the avian sarcoma virus src gene product. Proc Natl Acad Sci U S A. 1978 Apr;75(4):2021–2024. doi: 10.1073/pnas.75.4.2021. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Collett M. S., Purchio A. F., Erikson R. L. Avian sarcoma virus-transforming protein, pp60src shows protein kinase activity specific for tyrosine. Nature. 1980 May 15;285(5761):167–169. doi: 10.1038/285167a0. [DOI] [PubMed] [Google Scholar]
  4. Crouch T. H., Klee C. B. Positive cooperative binding of calcium to bovine brain calmodulin. Biochemistry. 1980 Aug 5;19(16):3692–3698. doi: 10.1021/bi00557a009. [DOI] [PubMed] [Google Scholar]
  5. Decker S., Lipmann F. Transport of D-glucose by membrane vesicles from normal and avian sarcoma virus-transformed chicken embryo fibroblasts. Proc Natl Acad Sci U S A. 1981 Sep;78(9):5358–5361. doi: 10.1073/pnas.78.9.5358. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Donner P., Bunte T., Owada M. K., Moelling K. Biochemical characterization of pp60src-associated protein kinase from avian sarcoma virus Schmidt-Ruppin strain. J Biol Chem. 1981 Aug 25;256(16):8786–8794. [PubMed] [Google Scholar]
  7. Fukami Y., Lipmann F. Purification of the Rous sarcoma virus src kinase by casein-agarose and tyrosine-agarose affinity chromatography. Proc Natl Acad Sci U S A. 1985 Jan;82(2):321–324. doi: 10.1073/pnas.82.2.321. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gerke V., Weber K. Identity of p36K phosphorylated upon Rous sarcoma virus transformation with a protein purified from brush borders; calcium-dependent binding to non-erythroid spectrin and F-actin. EMBO J. 1984 Jan;3(1):227–233. doi: 10.1002/j.1460-2075.1984.tb01789.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Glenney J. R., Jr, Glenney P., Osborn M., Weber K. An F-actin- and calmodulin-binding protein from isolated intestinal brush borders has a morphology related to spectrin. Cell. 1982 Apr;28(4):843–854. doi: 10.1016/0092-8674(82)90063-0. [DOI] [PubMed] [Google Scholar]
  10. Guild B. C., Erikson R. L., Strominger J. L. HLA-A2 and HLA-B7 antigens are phosphorylated in vitro by rous sarcoma virus kinase (pp60v-src) at a tyrosine residue encoded in a highly conserved exon of the intracellular domain. Proc Natl Acad Sci U S A. 1983 May;80(10):2894–2898. doi: 10.1073/pnas.80.10.2894. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hunter T., Sefton B. M. Transforming gene product of Rous sarcoma virus phosphorylates tyrosine. Proc Natl Acad Sci U S A. 1980 Mar;77(3):1311–1315. doi: 10.1073/pnas.77.3.1311. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kakiuchi S., Sobue K., Yamazaki R., Kambayashi J., Sakon M., Kosaki G. Lack of tissue specificity of calmodulin: a rapid and high-yield purification method. FEBS Lett. 1981 Apr 20;126(2):203–207. doi: 10.1016/0014-5793(81)80242-6. [DOI] [PubMed] [Google Scholar]
  13. Klee C. B., Crouch T. H., Richman P. G. Calmodulin. Annu Rev Biochem. 1980;49:489–515. doi: 10.1146/annurev.bi.49.070180.002421. [DOI] [PubMed] [Google Scholar]
  14. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  15. Lazarides E., Nelson W. J. Expression of spectrin in nonerythroid cells. Cell. 1982 Dec;31(3 Pt 2):505–508. doi: 10.1016/0092-8674(82)90306-3. [DOI] [PubMed] [Google Scholar]
  16. Levinson A. D., Oppermann H., Levintow L., Varmus H. E., Bishop J. M. Evidence that the transforming gene of avian sarcoma virus encodes a protein kinase associated with a phosphoprotein. Cell. 1978 Oct;15(2):561–572. doi: 10.1016/0092-8674(78)90024-7. [DOI] [PubMed] [Google Scholar]
  17. MITCHELL H. K., LUNAN K. D. TYROSINE-O-PHOSPHATE IN DROSOPHILA. Arch Biochem Biophys. 1964 Jul 20;106:219–222. doi: 10.1016/0003-9861(64)90179-1. [DOI] [PubMed] [Google Scholar]
  18. Ogawa Y., Tanokura M. Calcium binding to calmodulin: effects of ionic strength, Mg2+, pH and temperature. J Biochem. 1984 Jan;95(1):19–28. doi: 10.1093/oxfordjournals.jbchem.a134584. [DOI] [PubMed] [Google Scholar]
  19. Plancke Y. D., Lazarides E. Evidence for a phosphorylated form of calmodulin in chicken brain and muscle. Mol Cell Biol. 1983 Aug;3(8):1412–1420. doi: 10.1128/mcb.3.8.1412. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Purchio A. F., Erikson E., Brugge J. S., Erikson R. L. Identification of a polypeptide encoded by the avian sarcoma virus src gene. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1567–1571. doi: 10.1073/pnas.75.3.1567. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Putkey J. A., Ts'ui K. F., Tanaka T., Lagacé L., Stein J. P., Lai E. C., Means A. R. Chicken calmodulin genes. A species comparison of cDNA sequences and isolation of a genomic clone. J Biol Chem. 1983 Oct 10;258(19):11864–11870. [PubMed] [Google Scholar]
  22. 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]
  23. Sefton B. M., Hunter T., Ball E. H., Singer S. J. Vinculin: a cytoskeletal target of the transforming protein of Rous sarcoma virus. Cell. 1981 Apr;24(1):165–174. doi: 10.1016/0092-8674(81)90512-2. [DOI] [PubMed] [Google Scholar]
  24. Shinozawa T., Hayashi F. [Improved method of the synthesis of super-hot (gamma-32P)ATP]. Seikagaku. 1983 Sep;55(9):1099–1101. [PubMed] [Google Scholar]
  25. Trowsdale J., Lee J., Carey J., Grosveld F., Bodmer J., Bodmer W. Sequences related to HLA-DR alpha chain on human chromosome 6: restriction enzyme polymorphism detected with DC alpha chain probes. Proc Natl Acad Sci U S A. 1983 Apr;80(7):1972–1976. doi: 10.1073/pnas.80.7.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Tse-Dinh Y. C., Wong T. W., Goldberg A. R. Virus- and cell-encoded tyrosine protein kinases inactivate DNA topoisomerases in vitro. Nature. 1984 Dec 20;312(5996):785–786. doi: 10.1038/312785a0. [DOI] [PubMed] [Google Scholar]
  27. Van Eldik L. J., Burgess W. H. Analytical subcellular distribution of calmodulin and calmodulin-binding proteins in normal and virus-transformed fibroblasts. J Biol Chem. 1983 Apr 10;258(7):4539–4547. [PubMed] [Google Scholar]
  28. Yazawa M., Sakuma M., Yagi K. Calmodulins from muscles of marine invertebrates, scallop and sea anemone. J Biochem. 1980 May;87(5):1313–1320. doi: 10.1093/oxfordjournals.jbchem.a132869. [DOI] [PubMed] [Google Scholar]
  29. Yu K. T., Werth D. K., Pastan I. H., Czech M. P. src kinase catalyzes the phosphorylation and activation of the insulin receptor kinase. J Biol Chem. 1985 May 10;260(9):5838–5846. [PubMed] [Google Scholar]

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