Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1987 Dec;84(24):9064–9068. doi: 10.1073/pnas.84.24.9064

A conserved domain regulates interactions of the v-fps protein-tyrosine kinase with the host cell.

J E DeClue 1, I Sadowski 1, G S Martin 1, T Pawson 1
PMCID: PMC299692  PMID: 3480531

Abstract

All cytoplasmic protein-tyrosine kinases (PTKs) share a noncatalytic domain, termed SH2, which comprises approximately 100 residues located immediately N-terminal to the kinase domain. A linker in the AX9m mutant of Fujinami avian sarcoma virus (FSV) introduces a dipeptide insertion into the SH2 domain of the P130gag-fps PTK, which abolishes its ability to transform Rat-2 cells. However, at 36 degrees C AX9m FSV elicits focus formation and agar colony formation in infected chicken embryo fibroblasts (CEF) with single hit kinetics. At 41.5 degrees C AX9m FSV is nontransforming for CEF, and the mutant is therefore both host and temperature dependent for transforming activity. Both in vitro and in vivo, the specific kinase activity of AX9m FSV P130gag-fps, measured by autophosphorylation and phosphorylation of exogenous substrates, correlated with transforming activity. The consequences of the AX9m mutation for enzymatic function and transforming activity therefore depend on the cellular environment in which the altered v-fps protein is expressed. We conclude that the SH2 domain directs the interaction of the P130gag-fps catalytic domain with cellular proteins such as substrates for phosphorylation or regulators of kinase activity important for its transforming ability.

Full text

PDF
9064

Images in this article

9065Image
on p.9065
9066Image
on p.9066
9067Image
on p.9067

Selected References

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

  1. Bryant D. L., Parsons J. T. Amino acid alterations within a highly conserved region of the Rous sarcoma virus src gene product pp60src inactivate tyrosine protein kinase activity. Mol Cell Biol. 1984 May;4(5):862–866. doi: 10.1128/mcb.4.5.862. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. 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]
  3. DeClue J. E., Martin G. S. Phosphorylation of talin at tyrosine in Rous sarcoma virus-transformed cells. Mol Cell Biol. 1987 Jan;7(1):371–378. doi: 10.1128/mcb.7.1.371. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Feldman R. A., Wang E., Hanafusa H. Cytoplasmic localization of the transforming protein of Fujinami sarcoma virus: salt-sensitive association with subcellular components. J Virol. 1983 Feb;45(2):782–791. doi: 10.1128/jvi.45.2.782-791.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Gilmore T., DeClue J. E., Martin G. S. Protein phosphorylation at tyrosine is induced by the v-erbB gene product in vivo and in vitro. Cell. 1985 Mar;40(3):609–618. doi: 10.1016/0092-8674(85)90209-0. [DOI] [PubMed] [Google Scholar]
  6. Guyden J. C., Martin G. S. Transformation parameters of chick embryo fibroblasts transformed by Fujinami, PRCII, PRCII-p, and Y73 avian sarcoma viruses. Virology. 1982 Oct 15;122(1):71–83. doi: 10.1016/0042-6822(82)90378-6. [DOI] [PubMed] [Google Scholar]
  7. Hunter T., Cooper J. A. Protein-tyrosine kinases. Annu Rev Biochem. 1985;54:897–930. doi: 10.1146/annurev.bi.54.070185.004341. [DOI] [PubMed] [Google Scholar]
  8. Ingman-Baker J., Hinze E., Levy J. G., Pawson T. Monoclonal antibodies to the transforming protein of Fujinami avian sarcoma virus discriminate between different fps-encoded proteins. J Virol. 1984 May;50(2):572–578. doi: 10.1128/jvi.50.2.572-578.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Jakobovits E. B., Majors J. E., Varmus H. E. Hormonal regulation of the Rous sarcoma virus src gene via a heterologous promoter defines a threshold dose for cellular transformation. Cell. 1984 Oct;38(3):757–765. doi: 10.1016/0092-8674(84)90271-x. [DOI] [PubMed] [Google Scholar]
  10. Jove R., Mayer B. J., Iba H., Laugier D., Poirier F., Calothy G., Hanafusa T., Hanafusa H. Genetic analysis of p60v-src domains involved in the induction of different cell transformation parameters. J Virol. 1986 Dec;60(3):840–848. doi: 10.1128/jvi.60.3.840-848.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kawai S., Duesberg P. H., Hanafusa H. Transformation-defective mutants of Rous sarcoma virus with src gene deletions of varying length. J Virol. 1977 Dec;24(3):910–914. doi: 10.1128/jvi.24.3.910-914.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Martin G. S., Radke K., Hughes S., Quintrell N., Bishop J. M., Varmus H. E. Mutants of Rous sarcoma virus with extensive deletions of the viral genome. Virology. 1979 Jul 30;96(2):530–546. doi: 10.1016/0042-6822(79)90109-0. [DOI] [PubMed] [Google Scholar]
  13. Meckling-Hansen K., Nelson R., Branton P., Pawson T. Enzymatic activation of Fujinami sarcoma virus gag-fps transforming proteins by autophosphorylation at tyrosine. EMBO J. 1987 Mar;6(3):659–666. doi: 10.1002/j.1460-2075.1987.tb04805.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Pawson T., Guyden J., Kung T. H., Radke K., Gilmore T., Martin G. S. A strain of Fujinami sarcoma virus which is temperature-sensitive in protein phosphorylation and cellular transformation. Cell. 1980 Dec;22(3):767–775. doi: 10.1016/0092-8674(80)90553-x. [DOI] [PubMed] [Google Scholar]
  15. Radke K., Carter V. C., Moss P., Dehazya P., Schliwa M., Martin G. S. Membrane association of a 36,000-dalton substrate for tyrosine phosphorylation in chicken embryo fibroblasts transformed by avian sarcoma viruses. J Cell Biol. 1983 Nov;97(5 Pt 1):1601–1611. doi: 10.1083/jcb.97.5.1601. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Radke K., Martin G. S. Transformation by Rous sarcoma virus: effects of src gene expression on the synthesis and phosphorylation of cellular polypeptides. Proc Natl Acad Sci U S A. 1979 Oct;76(10):5212–5216. doi: 10.1073/pnas.76.10.5212. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Raymond V. W., Parsons J. T. Identification of an amino terminal domain required for the transforming activity of the Rous sarcoma virus src protein. Virology. 1987 Oct;160(2):400–410. doi: 10.1016/0042-6822(87)90011-0. [DOI] [PubMed] [Google Scholar]
  18. Sadowski I., Pawson T. Catalytic and non-catalytic domains of the Fujinami sarcoma virus P130gag-fps protein-tyrosine kinase distinguished by the expression of v-fps polypeptides in Escherichia coli. Oncogene. 1987 May;1(2):181–191. [PubMed] [Google Scholar]
  19. Sadowski I., Stone J. C., Pawson T. A noncatalytic domain conserved among cytoplasmic protein-tyrosine kinases modifies the kinase function and transforming activity of Fujinami sarcoma virus P130gag-fps. Mol Cell Biol. 1986 Dec;6(12):4396–4408. doi: 10.1128/mcb.6.12.4396. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Steimer K. S., Boettiger D. Complementation rescue of Rous sarcoma virus from transformed mammalian cells by polyethylene glycol-mediated cell fusion. J Virol. 1977 Jul;23(1):133–141. doi: 10.1128/jvi.23.1.133-141.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Stone J. C., Atkinson T., Smith M., Pawson T. Identification of functional regions in the transforming protein of Fujinami sarcoma virus by in-phase insertion mutagenesis. Cell. 1984 Jun;37(2):549–558. doi: 10.1016/0092-8674(84)90385-4. [DOI] [PubMed] [Google Scholar]
  22. Varmus H. E., Quintrell N., Wyke J. Revertants of an ASV-transformed rat cell line have lost the complete provius or sustained mutations in src. Virology. 1981 Jan 15;108(1):28–46. doi: 10.1016/0042-6822(81)90525-0. [DOI] [PubMed] [Google Scholar]
  23. Weinmaster G., Hinze E., Pawson T. Mapping of multiple phosphorylation sites within the structural and catalytic domains of the Fujinami avian sarcoma virus transforming protein. J Virol. 1983 Apr;46(1):29–41. doi: 10.1128/jvi.46.1.29-41.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Weinmaster G., Zoller M. J., Pawson T. A lysine in the ATP-binding site of P130gag-fps is essential for protein-tyrosine kinase activity. EMBO J. 1986 Jan;5(1):69–76. doi: 10.1002/j.1460-2075.1986.tb04179.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Weinmaster G., Zoller M. J., Smith M., Hinze E., Pawson T. Mutagenesis of Fujinami sarcoma virus: evidence that tyrosine phosphorylation of P130gag-fps modulates its biological activity. Cell. 1984 Jun;37(2):559–568. doi: 10.1016/0092-8674(84)90386-6. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES