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. 1984 Sep 1;99(3):788–795. doi: 10.1083/jcb.99.3.788

Tyrosyl kinases acquired from anchorage-independent cells by a membrane- enveloped virus

PMCID: PMC2113403  PMID: 6206078

Abstract

Tyrosyl kinase activity in vesicular stomatitis virus (VSV) acquired from host cells that differ in morphology was investigated. VSV grown in baby hamster kidney (BHK) cells with rounded morphology and a high efficiency of colony formation in soft agar (Rous sarcoma virus [RSV]- transformed and suspension BHK cells) was compared with VSV grown in BHK cells with a flattened morphology and lower efficiency of colony formation in soft agar (RSV-infected revertant and control BHK cells). Tyrosyl kinase activity measured with the substrates angiotensin II peptide or casein was found at 7-10-fold higher levels in virus released from the anchorage-independent BHK cells. Most of the VSV- associated tyrosyl kinases acquired from the RSV-transformed BHK cells reacted with antiserum to pp60src, whereas the activity acquired from the suspension BHK cells was unaffected by anti-src serum. The overall levels of tyrosyl kinase in subcellular fractions of the host BHK cells were also measured. Like the VSV released from them, the RSV- transformed cell extracts contained high levels. The suspension cells, however, contained the same low levels of tyrosyl kinase as was found in the control BHK cell extracts. Therefore, tyrosyl kinase was concentrated and acquired by VSV from the anchorage-independent suspension BHK cells. VSV-associated protein kinases acquired from other cell types followed a similar pattern. Tyrosyl kinase levels were high in VSV released from suspension cultures (Chinese hamster ovary and HeLa) and from virally transformed cells (Kirsten murine sarcoma virus-transformed rat kidney cells) and low in VSV released from an anchorage-dependent primary cell culture (chick embryo fibroblasts).

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Selected References

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  1. Alonso F. V., Compans R. W. Differential effect of monensin on enveloped viruses that form at distinct plasma membrane domains. J Cell Biol. 1981 Jun;89(3):700–705. doi: 10.1083/jcb.89.3.700. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bishop J. M. Cellular oncogenes and retroviruses. Annu Rev Biochem. 1983;52:301–354. doi: 10.1146/annurev.bi.52.070183.001505. [DOI] [PubMed] [Google Scholar]
  3. Boss M. A., Dreyfuss G., Baltimore D. Localization of the Abelson murine leukemia virus protein in a detergent-insoluble subcellular matrix: architecture of the protein. J Virol. 1981 Nov;40(2):472–481. doi: 10.1128/jvi.40.2.472-481.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bouck N., di Mayorca G. Chemical carcinogens transform BHK cells by inducing a recessive mutation. Mol Cell Biol. 1982 Feb;2(2):97–105. doi: 10.1128/mcb.2.2.97. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Burr J. G., Dreyfuss G., Penman S., Buchanan J. M. Association of the src gene product of Rous sarcoma virus with cytoskeletal structures of chicken embryo fibroblasts. Proc Natl Acad Sci U S A. 1980 Jun;77(6):3484–3488. doi: 10.1073/pnas.77.6.3484. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. CAPSTICK P. B., TELLING R. C., CHAPMAN W. G., STEWART D. L. Growth of a cloned strain of hamster kidney cells in suspended cultures and their susceptibility to the virus of foot-and-mouth disease. Nature. 1962 Sep 22;195:1163–1164. doi: 10.1038/1951163a0. [DOI] [PubMed] [Google Scholar]
  7. Choppin P. W., Compans R. W. Phenotypic mixing of envelope proteins of the parainfluenza virus SV5 and vesicular stomatitis virus. J Virol. 1970 May;5(5):609–616. doi: 10.1128/jvi.5.5.609-616.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Clinton G. M., Guerina N. G., Guo H. Y., Huang A. S. Host-dependent phosphorylation and kinase activity associated with vesicular stomatitis virus. J Biol Chem. 1982 Mar 25;257(6):3313–3319. [PubMed] [Google Scholar]
  9. Clinton G. M., Huang A. S. Distribution of phosphoserine, phosphothreonine and phosphotyrosine in proteins of vesicular stomatitis virus. Virology. 1981 Jan 30;108(2):510–514. doi: 10.1016/0042-6822(81)90459-1. [DOI] [PubMed] [Google Scholar]
  10. Clinton G. M., Roskoski R., Jr Tyrosyl and cyclic AMP-dependent protein kinase activities in BHK cells that express viral pp60src. Mol Cell Biol. 1984 May;4(5):973–977. doi: 10.1128/mcb.4.5.973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. 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]
  12. 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]
  13. 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]
  14. 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]
  15. Cooper J. A., Hunter T. Similarities and differences between the effects of epidermal growth factor and Rous sarcoma virus. J Cell Biol. 1981 Dec;91(3 Pt 1):878–883. doi: 10.1083/jcb.91.3.878. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Courtneidge S. A., Levinson A. D., Bishop J. M. The protein encoded by the transforming gene of avian sarcoma virus (pp60src) and a homologous protein in normal cells (pp60proto-src) are associated with the plasma membrane. Proc Natl Acad Sci U S A. 1980 Jul;77(7):3783–3787. doi: 10.1073/pnas.77.7.3783. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Deng C. T., Stehelin D., Bishop J. M., Varmus H. E. Characteristics of virus-specific RNA in avian sarcoma virus-transformed BHK-21 cells and revertants. Virology. 1977 Jan;76(1):313–330. doi: 10.1016/0042-6822(77)90305-1. [DOI] [PubMed] [Google Scholar]
  18. Di Mayorca G., Greenblatt M., Trauthen T., Soller A., Giordano R. Malignant transformation of BHK21 clone 13 cells in vitro by nitrosamines--a conditional state. Proc Natl Acad Sci U S A. 1973 Jan;70(1):46–49. doi: 10.1073/pnas.70.1.46. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. 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]
  20. 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]
  21. HAM R. G. Clonal growth of diploid Chinese hamster cells in a synthetic medium supplemented with purified protein fractions. Exp Cell Res. 1962 Dec;28:489–500. doi: 10.1016/0014-4827(62)90253-7. [DOI] [PubMed] [Google Scholar]
  22. Huang A. S., Palma E. L., Hewlett N., Roizman B. Pseudotype formation between enveloped RNA and DNA viruses. Nature. 1974 Dec 20;252(5485):743–745. doi: 10.1038/252743a0. [DOI] [PubMed] [Google Scholar]
  23. 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]
  24. Hynes R. Phosphorylation of vinculin by pp60src: what might it mean? Cell. 1982 Mar;28(3):437–438. doi: 10.1016/0092-8674(82)90196-9. [DOI] [PubMed] [Google Scholar]
  25. Little L. M., Zavada J., Der C. J., Huang A. S. Identity of HeLa cell determinants acquired by vesicular stomatitis virus with a tumor antigen. Science. 1983 Jun 3;220(4601):1069–1071. doi: 10.1126/science.6302845. [DOI] [PubMed] [Google Scholar]
  26. Lodish H. F., Porter M. Specific incorporation of host cell surface proteins into budding vesicular stomatitis virus particles. Cell. 1980 Jan;19(1):161–169. doi: 10.1016/0092-8674(80)90397-9. [DOI] [PubMed] [Google Scholar]
  27. Lopez V., Stevens T., Lindquist R. N. Vanadium ion inhibition of alkaline phosphatase-catalyzed phosphate ester hydrolysis. Arch Biochem Biophys. 1976 Jul;175(1):31–38. doi: 10.1016/0003-9861(76)90482-3. [DOI] [PubMed] [Google Scholar]
  28. MACPHERSON I., MONTAGNIER L. AGAR SUSPENSION CULTURE FOR THE SELECTIVE ASSAY OF CELLS TRANSFORMED BY POLYOMA VIRUS. Virology. 1964 Jun;23:291–294. doi: 10.1016/0042-6822(64)90301-0. [DOI] [PubMed] [Google Scholar]
  29. McLIMANS W. F., DAVIS E. V., GLOVER F. L., RAKE G. W. The submerged culture of mammalian cells; the spinner culture. J Immunol. 1957 Nov;79(5):428–433. [PubMed] [Google Scholar]
  30. 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]
  31. Owada M. K., Donner P., Scott A., Moelling K. Isolation of an avian sarcoma virus-specific protein kinase from virus particles. Virology. 1981 Apr 30;110(2):333–343. doi: 10.1016/0042-6822(81)90064-7. [DOI] [PubMed] [Google Scholar]
  32. Petruzzelli L. M., Ganguly S., Smith C. J., Cobb M. H., Rubin C. S., Rosen O. M. Insulin activates a tyrosine-specific protein kinase in extracts of 3T3-L1 adipocytes and human placenta. Proc Natl Acad Sci U S A. 1982 Nov;79(22):6792–6796. doi: 10.1073/pnas.79.22.6792. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Quintrell N., Hughes S. H., Varmus H. E., Bishop J. M. Structure of viral DNA and RNA in mammalian cells infected with avian sarcoma virus. J Mol Biol. 1980 Nov 15;143(4):363–393. doi: 10.1016/0022-2836(80)90218-1. [DOI] [PubMed] [Google Scholar]
  34. Rodriguez Boulan E., Pendergast M. Polarized distribution of viral envelope proteins in the plasma membrane of infected epithelial cells. Cell. 1980 May;20(1):45–54. doi: 10.1016/0092-8674(80)90233-0. [DOI] [PubMed] [Google Scholar]
  35. Rodriguez Boulan E., Sabatini D. D. Asymmetric budding of viruses in epithelial monlayers: a model system for study of epithelial polarity. Proc Natl Acad Sci U S A. 1978 Oct;75(10):5071–5075. doi: 10.1073/pnas.75.10.5071. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. 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]
  37. 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]
  38. Roth M. G., Compans R. W. Antibody-resistant spread of vesicular stomatitis virus infection in cell lines of epithelial origin. J Virol. 1980 Aug;35(2):547–550. doi: 10.1128/jvi.35.2.547-550.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Roth R. A., Cassell D. J. Insulin receptor: evidence that it is a protein kinase. Science. 1983 Jan 21;219(4582):299–301. doi: 10.1126/science.6849137. [DOI] [PubMed] [Google Scholar]
  40. STOKER M., MACPHERSON I. SYRIAN HAMSTER FIBROBLAST CELL LINE BHK21 AND ITS DERIVATIVES. Nature. 1964 Sep 26;203:1355–1357. doi: 10.1038/2031355a0. [DOI] [PubMed] [Google Scholar]
  41. 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]
  42. 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]
  43. Shih T. Y., Papageorge A. G., Stokes P. E., Weeks M. O., Scolnick E. M. Guanine nucleotide-binding and autophosphorylating activities associated with the p21src protein of Harvey murine sarcoma virus. Nature. 1980 Oct 23;287(5784):686–691. doi: 10.1038/287686a0. [DOI] [PubMed] [Google Scholar]
  44. Shih T. Y., Weeks M. O., Young H. A., Scolnick E. M. p21 of Kirsten murine sarcoma virus is thermolabile in a viral mutant temperature sensitive for the maintenance of transformation. J Virol. 1979 Aug;31(2):546–546. doi: 10.1128/jvi.31.2.546-546.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Shriver K., Rohrschneider L. Organization of pp60src and selected cytoskeletal proteins within adhesion plaques and junctions of Rous sarcoma virus-transformed rat cells. J Cell Biol. 1981 Jun;89(3):525–535. doi: 10.1083/jcb.89.3.525. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Stadtmauer L. A., Rosen O. M. Phosphorylation of exogenous substrates by the insulin receptor-associated protein kinase. J Biol Chem. 1983 Jun 10;258(11):6682–6685. [PubMed] [Google Scholar]
  47. Stiles C. D. The molecular biology of platelet-derived growth factor. Cell. 1983 Jul;33(3):653–655. doi: 10.1016/0092-8674(83)90008-9. [DOI] [PubMed] [Google Scholar]
  48. 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]
  49. Witt J. J., Roskoski R., Jr Rapid protein kinase assay using phosphocellulose-paper absorption. Anal Biochem. 1975 May 26;66(1):253–258. doi: 10.1016/0003-2697(75)90743-5. [DOI] [PubMed] [Google Scholar]
  50. Wong T. W., Goldberg A. R. In vitro phosphorylation of angiotensin analogs by tyrosyl protein kinases. J Biol Chem. 1983 Jan 25;258(2):1022–1025. [PubMed] [Google Scholar]
  51. Wong T. W., Goldberg A. R. Tyrosyl protein kinases in normal rat liver: identification and partial characterization. Proc Natl Acad Sci U S A. 1983 May;80(9):2529–2533. doi: 10.1073/pnas.80.9.2529. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Závada J. VSV pseudotype particles with the coat of avian myeloblastosis virus. Nat New Biol. 1972 Nov 22;240(99):122–124. doi: 10.1038/newbio240122a0. [DOI] [PubMed] [Google Scholar]

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