Skip to main content
NCBI home page
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1984 Aug;4(8):1508–1514. doi: 10.1128/mcb.4.8.1508

Functional domains of the pp60v-src protein as revealed by analysis of temperature-sensitive Rous sarcoma virus mutants.

A W Stoker, P J Enrietto, J A Wyke
PMCID: PMC368941  PMID: 6092915

Abstract

Four temperature-sensitive (ts) Rous sarcoma virus src gene mutants with lesions in different parts of the gene represent three classes of alteration in pp60src. These classes are composed of mutants with (i) heat-labile protein kinase activities both in vitro and in vivo (tsLA27 and tsLA29), (ii) heat-labile kinases in vivo but not in vitro (tsLA33), and (iii) neither in vivo nor in vitro heat-labile kinases (tsLA32). The latter class indicates the existence of structural or functional pp60src domains that are required for transformation but do not grossly affect tyrosine kinase activity.

Full text

PDF
1508

Images in this article

1509Image
on p.1509
1510Image
on p.1510
1511Image
on p.1511
1512Image
on p.1512
1512Image
on p.1512

Selected References

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

  1. Amini S., Kaji A. Association of pp36, a phosphorylated form of the presumed target protein for the src protein of Rous sarcoma virus, with the membrane of chicken cells transformed by Rous sarcoma virus. Proc Natl Acad Sci U S A. 1983 Feb;80(4):960–964. doi: 10.1073/pnas.80.4.960. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Anderson D. D., Beckmann R. P., Harms E. H., Nakamura K., Weber M. J. Biological properties of "partial" transformation mutants of Rous sarcoma virus and characterization of their pp60src kinase. J Virol. 1981 Jan;37(1):445–458. doi: 10.1128/jvi.37.1.445-458.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Anderson D. D., Salter D. W., Rohrschneider L. R., Weber M. J. Genetic and biochemical approaches to analyzing transformation by Rous sarcoma virus. Cold Spring Harb Symp Quant Biol. 1980;44(Pt 2):1031–1041. doi: 10.1101/sqb.1980.044.01.111. [DOI] [PubMed] [Google Scholar]
  4. Becker D., Kurth R., Critchley D., Friis R., Bauer H. Distinguishable transformation-defective phenotypes among temperature-sensitive mutants of Rous sarcoma virus. J Virol. 1977 Mar;21(3):1042–1055. doi: 10.1128/jvi.21.3.1042-1055.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Beemon K., Ryden T., McNelly E. A. Transformation by avian sarcoma viruses leads to phosphorylation of multiple cellular proteins on tyrosine residues. J Virol. 1982 May;42(2):742–747. doi: 10.1128/jvi.42.2.742-747.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Brugge J. S., Erikson E., Erikson R. L. The specific interaction of the Rous sarcoma virus transforming protein, pp60src, with two cellular proteins. Cell. 1981 Aug;25(2):363–372. doi: 10.1016/0092-8674(81)90055-6. [DOI] [PubMed] [Google Scholar]
  7. Brugge J. S., Steinbaugh P. J., Erikson R. L. Characterization of the avian sarcoma virus protein p60src. Virology. 1978 Nov;91(1):130–140. doi: 10.1016/0042-6822(78)90361-6. [DOI] [PubMed] [Google Scholar]
  8. Bryant D., Parsons J. T. Site-directed mutagenesis of the src gene of Rous sarcoma virus: construction and characterization of a deletion mutant temperature sensitive for transformation. J Virol. 1982 Nov;44(2):683–691. doi: 10.1128/jvi.44.2.683-691.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Cheng Y. S., Chen L. B. Detection of phosphotyrosine-containing 34,000-dalton protein in the framework of cells transformed with Rous sarcoma virus. Proc Natl Acad Sci U S A. 1981 Apr;78(4):2388–2392. doi: 10.1073/pnas.78.4.2388. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. 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]
  11. Collett M. S., Erikson E., Erikson R. L. Structural analysis of the avian sarcoma virus transforming protein: sites of phosphorylation. J Virol. 1979 Feb;29(2):770–781. doi: 10.1128/jvi.29.2.770-781.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. 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]
  14. 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]
  15. Cooper J. A., Hunter T. Discrete primary locations of a tyrosine-protein kinase and of three proteins that contain phosphotyrosine in virally transformed chick fibroblasts. J Cell Biol. 1982 Aug;94(2):287–296. doi: 10.1083/jcb.94.2.287. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. 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]
  17. 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]
  18. Cooper J., Nakamura K. D., Hunter T., Weber M. J. Phosphotyrosine-containing proteins and expression of transformation parameters in cells infected with partial transformation mutants of Rous sarcoma virus. J Virol. 1983 Apr;46(1):15–28. doi: 10.1128/jvi.46.1.15-28.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Courtneidge S. A., Bishop J. M. Transit of pp60v-src to the plasma membrane. Proc Natl Acad Sci U S A. 1982 Dec;79(23):7117–7121. doi: 10.1073/pnas.79.23.7117. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Courtneidge S., Ralston R., Alitalo K., Bishop J. M. Subcellular location of an abundant substrate (p36) for tyrosine-specific protein kinases. Mol Cell Biol. 1983 Mar;3(3):340–350. doi: 10.1128/mcb.3.3.340. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Cross F. R., Hanafusa H. Local mutagenesis of Rous sarcoma virus: the major sites of tyrosine and serine phosphorylation of pp60src are dispensable for transformation. Cell. 1983 Sep;34(2):597–607. doi: 10.1016/0092-8674(83)90392-6. [DOI] [PubMed] [Google Scholar]
  22. Enrietto P. J., Payne L. N., Wyke J. A. Analysis of the pathogenicity of transformation defective partial deletion mutants of avian sarcoma virus: characterization of recovered viruses which encode novel src specific proteins. Virology. 1983 Jun;127(2):397–411. doi: 10.1016/0042-6822(83)90153-8. [DOI] [PubMed] [Google Scholar]
  23. Erikson E., Collett M. S., Erikson R. L. In vitro synthesis of a functional avian sarcoma virus transforming-gene product. Nature. 1978 Aug 31;274(5674):919–921. doi: 10.1038/274919a0. [DOI] [PubMed] [Google Scholar]
  24. 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]
  25. Erikson R. L., Collett M. S., Erikson E., Purchio A. F. Evidence that the avian sarcoma virus transforming gene product is a cyclic AMP-independent protein kinase. Proc Natl Acad Sci U S A. 1979 Dec;76(12):6260–6264. doi: 10.1073/pnas.76.12.6260. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Fincham V. J., Chiswell D. J., Wyke J. A. Mapping of nonconditional and conditional mutants in the src gene of Prague strain Rous sarcoma virus. Virology. 1982 Jan 15;116(1):72–83. doi: 10.1016/0042-6822(82)90404-4. [DOI] [PubMed] [Google Scholar]
  27. Friis R. R., Schwarz R. T., Schmidt M. F. Phenotypes of Rous sarcoma virus-transformed fibroblasts: an argument for a multifunctional Src gene product. Med Microbiol Immunol. 1977;164(1-3):155–165. doi: 10.1007/BF02121311. [DOI] [PubMed] [Google Scholar]
  28. Garber E. A., Krueger J. G., Hanafusa H., Goldberg A. R. Temperature-sensitive membrane association of pp60src in tsNY68-infected cells correlates with increased tyrosine phosphorylation of membrane-associated proteins. Virology. 1983 Apr 15;126(1):73–86. doi: 10.1016/0042-6822(83)90462-2. [DOI] [PubMed] [Google Scholar]
  29. Gilmore T. D., Radke K., Martin G. S. Tyrosine phosphorylation of a 50K cellular polypeptide associated with the Rous sarcoma virus transforming protein pp60src. Mol Cell Biol. 1982 Feb;2(2):199–206. doi: 10.1128/mcb.2.2.199. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Goldberg A. R., Krueger J. G., Wang E. Localization and characterization of the src-gene product of Rous sarcoma virus. Cold Spring Harb Symp Quant Biol. 1980;44(Pt 2):991–1005. doi: 10.1101/sqb.1980.044.01.107. [DOI] [PubMed] [Google Scholar]
  31. 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]
  32. Kawai S., Hanafusa H. The effects of reciprocal changes in temperature on the transformed state of cells infected with a rous sarcoma virus mutant. Virology. 1971 Nov;46(2):470–479. doi: 10.1016/0042-6822(71)90047-x. [DOI] [PubMed] [Google Scholar]
  33. Krueger J. G., Garber E. A., Goldberg A. R., Hanafusa H. Changes in amino-terminal sequences of pp60src lead to decreased membrane association and decreased in vivo tumorigenicity. Cell. 1982 Apr;28(4):889–896. doi: 10.1016/0092-8674(82)90068-x. [DOI] [PubMed] [Google Scholar]
  34. Levinson A. D., Courtneidge S. A., Bishop J. M. Structural and functional domains of the Rous sarcoma virus transforming protein (pp60src). Proc Natl Acad Sci U S A. 1981 Mar;78(3):1624–1628. doi: 10.1073/pnas.78.3.1624. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. 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]
  36. Mardon G., Varmus H. E. Frameshift and intragenic suppressor mutations in a Rous sarcoma provirus suggest src encodes two proteins. Cell. 1983 Mar;32(3):871–879. doi: 10.1016/0092-8674(83)90072-7. [DOI] [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., Weber M. J. Phosphorylation of a 36,000 Mr cellular protein in cells infected with partial transformation mutants of rous sarcoma virus. Mol Cell Biol. 1982 Feb;2(2):147–153. doi: 10.1128/mcb.2.2.147. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Nigg E. A., Cooper J. A., Hunter T. Immunofluorescent localization of a 39,000-dalton substrate of tyrosine protein kinases to the cytoplasmic surface of the plasma membrane. J Cell Biol. 1983 Jun;96(6):1601–1609. doi: 10.1083/jcb.96.6.1601. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Oppermann H., Levinson A. D., Levintow L., Varmus H. E., Bishop J. M., Kawai S. Two cellular proteins that immunoprecipitate with the transforming protein of Rous sarcoma virus. Virology. 1981 Sep;113(2):736–751. doi: 10.1016/0042-6822(81)90202-6. [DOI] [PubMed] [Google Scholar]
  41. Oppermann H., Levinson A. D., Varmus H. E., Levintow L., Bishop J. M. Uninfected vertebrate cells contain a protein that is closely related to the product of the avian sarcoma virus transforming gene (src). Proc Natl Acad Sci U S A. 1979 Apr;76(4):1804–1808. doi: 10.1073/pnas.76.4.1804. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Parry G., Bartholomew J. C., Bissell M. J. Role of src gene in growth regulation of Rous sarcoma virus-infected chicken embryo fibroblasts. Nature. 1980 Dec 25;288(5792):720–722. doi: 10.1038/288720a0. [DOI] [PubMed] [Google Scholar]
  43. Pastan I., Jay G., Richert N., Davies P. J., Willingham M. Nature and intracellular location of the product of the src gene of avian sarcoma virus. Cold Spring Harb Symp Quant Biol. 1980;44(Pt 2):1023–1029. doi: 10.1101/sqb.1980.044.01.110. [DOI] [PubMed] [Google Scholar]
  44. 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]
  45. Purchio A. F. Evidence the pp60src, the product of the Rous sarcoma virus src gene, undergoes autophosphorylation. J Virol. 1982 Jan;41(1):1–7. doi: 10.1128/jvi.41.1.1-7.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. 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]
  47. 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]
  48. Rohrschneider L. R. Adhesion plaques of Rous sarcoma virus-transformed cells contain the src gene product. Proc Natl Acad Sci U S A. 1980 Jun;77(6):3514–3518. doi: 10.1073/pnas.77.6.3514. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Rohrschneider L., Rosok M. J. Transformation parameters and pp60src localization in cells infected with partial transformation mutants of Rous sarcoma virus. Mol Cell Biol. 1983 Apr;3(4):731–746. doi: 10.1128/mcb.3.4.731. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Schwartz D. E., Tizard R., Gilbert W. Nucleotide sequence of Rous sarcoma virus. Cell. 1983 Mar;32(3):853–869. doi: 10.1016/0092-8674(83)90071-5. [DOI] [PubMed] [Google Scholar]
  51. 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]
  52. Sefton B. M., Hunter T., Beemon K., Eckhart W. Evidence that the phosphorylation of tyrosine is essential for cellular transformation by Rous sarcoma virus. Cell. 1980 Jul;20(3):807–816. doi: 10.1016/0092-8674(80)90327-x. [DOI] [PubMed] [Google Scholar]
  53. Sefton B. M., Hunter T., Beemon K. Temperature-sensitive transformation by Rous sarcoma virus and temperature-sensitive protein kinase activity. J Virol. 1980 Jan;33(1):220–229. doi: 10.1128/jvi.33.1.220-229.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Sefton B. M., Hunter T., Cooper J. A. Some lymphoid cell lines transformed by Abelson murine leukemia virus lack a major 36,000-dalton tyrosine protein kinase substrate. Mol Cell Biol. 1983 Jan;3(1):56–63. doi: 10.1128/mcb.3.1.56. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Smart J. E., Oppermann H., Czernilofsky A. P., Purchio A. F., Erikson R. L., Bishop J. M. Characterization of sites for tyrosine phosphorylation in the transforming protein of Rous sarcoma virus (pp60v-src) and its normal cellular homologue (pp60c-src). Proc Natl Acad Sci U S A. 1981 Oct;78(10):6013–6017. doi: 10.1073/pnas.78.10.6013. [DOI] [PMC free article] [PubMed] [Google Scholar]
  56. Snyder M. A., Bishop J. M., Colby W. W., Levinson A. D. Phosphorylation of tyrosine-416 is not required for the transforming properties and kinase activity of pp60v-src. Cell. 1983 Mar;32(3):891–901. doi: 10.1016/0092-8674(83)90074-0. [DOI] [PubMed] [Google Scholar]
  57. Tamura T., Bauer H., Birr C., Pipkorn R. Antibodies against synthetic peptides as a tool for functional analysis of the transforming protein pp60src. Cell. 1983 Sep;34(2):587–596. doi: 10.1016/0092-8674(83)90391-4. [DOI] [PubMed] [Google Scholar]
  58. Tato F., Beamand J. A., Wyke J. A. A mutant of Rous sarcoma virus with a thermolabile defect in the virus envelope. Virology. 1978 Jul 1;88(1):71–81. doi: 10.1016/0042-6822(78)90111-3. [DOI] [PubMed] [Google Scholar]
  59. Weber M. J., Friis R. R. Dissociation of transformation parameters using temperature-conditional mutants of Rous sarcoma virus. Cell. 1979 Jan;16(1):25–32. doi: 10.1016/0092-8674(79)90184-3. [DOI] [PubMed] [Google Scholar]
  60. Wyke J. A., Linial M. Temperature-sensitive avian sarcoma viruses: a physiological comparison of twenty mutants. Virology. 1973 May;53(1):152–161. doi: 10.1016/0042-6822(73)90474-1. [DOI] [PubMed] [Google Scholar]

Articles from Molecular and Cellular Biology are provided here courtesy of Taylor & Francis

RESOURCES