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. 1996 Jan;70(1):600–606. doi: 10.1128/jvi.70.1.600-606.1996

A herpesvirus saimiri membrane protein required for interleukin-2 independence forms a stable complex with p56lck.

T Lund 1, M M Medveczky 1, P J Neame 1, P G Medveczky 1
PMCID: PMC189852  PMID: 8523578

Abstract

ORF-2, a 32-kDa viral protein expressed by herpesvirus saimiri-transformed lymphocytes, is essential for transformation and is expressed on the plasma membrane of transformed cells. The current work now shows that most (approximately 80%) of ORF-2 resides in the cytoplasm, while only a small portion protrudes from the cell surface. Expressed as a glutathione S-transferase fusion protein, ORF-2 was found to interact with a 56-kDa cellular protein in untransformed, herpesvirus saimiri-transformed, and Jurkat lymphocytes. Microsequencing proved that this protein is the lymphocyte-specific tyrosine protein kinase p56lck. Two regions of ORF-2 were found to be required for p56lck interaction. Current evidence suggests that the interaction of ORF-2 with p56lck plays a key role in the specific transformation of T lymphocytes to an interleukin-2-independent phenotype.

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Selected References

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  1. Abraham K. M., Levin S. D., Marth J. D., Forbush K. A., Perlmutter R. M. Thymic tumorigenesis induced by overexpression of p56lck. Proc Natl Acad Sci U S A. 1991 May 1;88(9):3977–3981. doi: 10.1073/pnas.88.9.3977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Altschul S. F., Gish W., Miller W., Myers E. W., Lipman D. J. Basic local alignment search tool. J Mol Biol. 1990 Oct 5;215(3):403–410. doi: 10.1016/S0022-2836(05)80360-2. [DOI] [PubMed] [Google Scholar]
  3. Biesinger B., Müller-Fleckenstein I., Simmer B., Lang G., Wittmann S., Platzer E., Desrosiers R. C., Fleckenstein B. Stable growth transformation of human T lymphocytes by herpesvirus saimiri. Proc Natl Acad Sci U S A. 1992 Apr 1;89(7):3116–3119. doi: 10.1073/pnas.89.7.3116. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Biesinger B., Trimble J. J., Desrosiers R. C., Fleckenstein B. The divergence between two oncogenic Herpesvirus saimiri strains in a genomic region related to the transforming phenotype. Virology. 1990 Jun;176(2):505–514. doi: 10.1016/0042-6822(90)90020-r. [DOI] [PubMed] [Google Scholar]
  5. Biesinger B., Tsygankov A. Y., Fickenscher H., Emmrich F., Fleckenstein B., Bolen J. B., Bröker B. M. The product of the Herpesvirus saimiri open reading frame 1 (tip) interacts with T cell-specific kinase p56lck in transformed cells. J Biol Chem. 1995 Mar 3;270(9):4729–4734. doi: 10.1074/jbc.270.9.4729. [DOI] [PubMed] [Google Scholar]
  6. Bornkamm G. W., Delius H., Fleckenstein B., Werner F. J., Mulder C. Structure of Herpesvirus saimiri genomes: arrangement of heavy and light sequences in the M genome. J Virol. 1976 Jul;19(1):154–161. doi: 10.1128/jvi.19.1.154-161.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Burkhardt A. L., Bolen J. B., Kieff E., Longnecker R. An Epstein-Barr virus transformation-associated membrane protein interacts with src family tyrosine kinases. J Virol. 1992 Aug;66(8):5161–5167. doi: 10.1128/jvi.66.8.5161-5167.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cheung R. K., Dosch H. M. The tyrosine kinase lck is critically involved in the growth transformation of human B lymphocytes. J Biol Chem. 1991 May 15;266(14):8667–8670. [PubMed] [Google Scholar]
  9. Chou C. S., Medveczky M. M., Geck P., Vercelli D., Medveczky P. G. Expression of IL-2 and IL-4 in T lymphocytes transformed by herpesvirus saimiri. Virology. 1995 Apr 20;208(2):418–426. doi: 10.1006/viro.1995.1172. [DOI] [PubMed] [Google Scholar]
  10. Churcher Y., Moss S. E. Modulation of protein tyrosine phosphorylation during G1/S transition in activated human T-lymphoblasts. J Biol Chem. 1993 Dec 15;268(35):26144–26149. [PubMed] [Google Scholar]
  11. Cole S. R., Ashman L. K., Ey P. L. Biotinylation: an alternative to radioiodination for the identification of cell surface antigens in immunoprecipitates. Mol Immunol. 1987 Jul;24(7):699–705. doi: 10.1016/0161-5890(87)90051-4. [DOI] [PubMed] [Google Scholar]
  12. Courtneidge S. A., Smith A. E. Polyoma virus transforming protein associates with the product of the c-src cellular gene. Nature. 1983 Jun 2;303(5916):435–439. doi: 10.1038/303435a0. [DOI] [PubMed] [Google Scholar]
  13. Fernandez J., DeMott M., Atherton D., Mische S. M. Internal protein sequence analysis: enzymatic digestion for less than 10 micrograms of protein bound to polyvinylidene difluoride or nitrocellulose membranes. Anal Biochem. 1992 Mar;201(2):255–264. doi: 10.1016/0003-2697(92)90336-6. [DOI] [PubMed] [Google Scholar]
  14. Geck P., Whitaker S. A., Medveczky M. M., Last T. J., Medveczky P. G. Small RNA expression from the oncogenic region of a highly oncogenic strain of herpesvirus saimiri. Virus Genes. 1994 Jan;8(1):25–34. doi: 10.1007/BF01703599. [DOI] [PubMed] [Google Scholar]
  15. Geck P., Whitaker S. A., Medveczky M. M., Medveczky P. G. Expression of collagenlike sequences by a tumor virus, herpesvirus saimiri. J Virol. 1990 Jul;64(7):3509–3515. doi: 10.1128/jvi.64.7.3509-3515.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hatakeyama M., Kono T., Kobayashi N., Kawahara A., Levin S. D., Perlmutter R. M., Taniguchi T. Interaction of the IL-2 receptor with the src-family kinase p56lck: identification of novel intermolecular association. Science. 1991 Jun 14;252(5012):1523–1528. doi: 10.1126/science.2047859. [DOI] [PubMed] [Google Scholar]
  17. Horak I. D., Gress R. E., Lucas P. J., Horak E. M., Waldmann T. A., Bolen J. B. T-lymphocyte interleukin 2-dependent tyrosine protein kinase signal transduction involves the activation of p56lck. Proc Natl Acad Sci U S A. 1991 Mar 1;88(5):1996–2000. doi: 10.1073/pnas.88.5.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Horak I. D., Kawakami T., Gregory F., Robbins K. C., Bolen J. B. Association of p60fyn with middle tumor antigen in murine polyomavirus-transformed rat cells. J Virol. 1989 May;63(5):2343–2347. doi: 10.1128/jvi.63.5.2343-2347.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Hubert P., Bismuth G., Körner M., Debré P. HIV-1 glycoprotein gp120 disrupts CD4-p56lck/CD3-T cell receptor interactions and inhibits CD3 signaling. Eur J Immunol. 1995 May;25(5):1417–1425. doi: 10.1002/eji.1830250542. [DOI] [PubMed] [Google Scholar]
  20. Hurley T. R., Hyman R., Sefton B. M. Differential effects of expression of the CD45 tyrosine protein phosphatase on the tyrosine phosphorylation of the lck, fyn, and c-src tyrosine protein kinases. Mol Cell Biol. 1993 Mar;13(3):1651–1656. doi: 10.1128/mcb.13.3.1651. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Iwashima M., Irving B. A., van Oers N. S., Chan A. C., Weiss A. Sequential interactions of the TCR with two distinct cytoplasmic tyrosine kinases. Science. 1994 Feb 25;263(5150):1136–1139. doi: 10.1126/science.7509083. [DOI] [PubMed] [Google Scholar]
  22. Izquierdo Pastor M., Reif K., Cantrell D. The regulation and function of p21ras during T-cell activation and growth. Immunol Today. 1995 Mar;16(3):159–164. doi: 10.1016/0167-5699(95)80134-0. [DOI] [PubMed] [Google Scholar]
  23. Karnitz L., Sutor S. L., Torigoe T., Reed J. C., Bell M. P., McKean D. J., Leibson P. J., Abraham R. T. Effects of p56lck deficiency on the growth and cytolytic effector function of an interleukin-2-dependent cytotoxic T-cell line. Mol Cell Biol. 1992 Oct;12(10):4521–4530. doi: 10.1128/mcb.12.10.4521. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Kornbluth S., Sudol M., Hanafusa H. Association of the polyomavirus middle-T antigen with c-yes protein. Nature. 1987 Jan 8;325(7000):171–173. doi: 10.1038/325171a0. [DOI] [PubMed] [Google Scholar]
  25. Lund T., Medveczky M. M., Geck P., Medveczky P. G. A herpesvirus saimiri protein required for interleukin-2 independence is associated with membranes of transformed T cells. J Virol. 1995 Jul;69(7):4495–4499. doi: 10.1128/jvi.69.7.4495-4499.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Luo K., Sefton B. M. Activated lck tyrosine protein kinase stimulates antigen-independent interleukin-2 production in T cells. Mol Cell Biol. 1992 Oct;12(10):4724–4732. doi: 10.1128/mcb.12.10.4724. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Medveczky M. M., Geck P., Sullivan J. L., Serbousek D., Djeu J. Y., Medveczky P. G. IL-2 independent growth and cytotoxicity of herpesvirus saimiri-infected human CD8 cells and involvement of two open reading frame sequences of the virus. Virology. 1993 Oct;196(2):402–412. doi: 10.1006/viro.1993.1495. [DOI] [PubMed] [Google Scholar]
  28. Medveczky M. M., Geck P., Vassallo R., Medveczky P. G. Expression of the collagen-like putative oncoprotein of Herpesvirus saimiri in transformed T cells. Virus Genes. 1993 Dec;7(4):349–365. doi: 10.1007/BF01703391. [DOI] [PubMed] [Google Scholar]
  29. Medveczky M. M., Szomolanyi E., Hesselton R., DeGrand D., Geck P., Medveczky P. G. Herpesvirus saimiri strains from three DNA subgroups have different oncogenic potentials in New Zealand white rabbits. J Virol. 1989 Sep;63(9):3601–3611. doi: 10.1128/jvi.63.9.3601-3611.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Medveczky P., Szomolanyi E., Desrosiers R. C., Mulder C. Classification of herpesvirus saimiri into three groups based on extreme variation in a DNA region required for oncogenicity. J Virol. 1984 Dec;52(3):938–944. doi: 10.1128/jvi.52.3.938-944.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Pawson T. SH2 and SH3 domains in signal transduction. Adv Cancer Res. 1994;64:87–110. doi: 10.1016/s0065-230x(08)60835-0. [DOI] [PubMed] [Google Scholar]
  32. Ren R., Mayer B. J., Cicchetti P., Baltimore D. Identification of a ten-amino acid proline-rich SH3 binding site. Science. 1993 Feb 19;259(5098):1157–1161. doi: 10.1126/science.8438166. [DOI] [PubMed] [Google Scholar]
  33. Resh M. D. Myristylation and palmitylation of Src family members: the fats of the matter. Cell. 1994 Feb 11;76(3):411–413. doi: 10.1016/0092-8674(94)90104-x. [DOI] [PubMed] [Google Scholar]
  34. Rudd C. E., Trevillyan J. M., Dasgupta J. D., Wong L. L., Schlossman S. F. The CD4 receptor is complexed in detergent lysates to a protein-tyrosine kinase (pp58) from human T lymphocytes. Proc Natl Acad Sci U S A. 1988 Jul;85(14):5190–5194. doi: 10.1073/pnas.85.14.5190. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Straus D. B., Weiss A. Genetic evidence for the involvement of the lck tyrosine kinase in signal transduction through the T cell antigen receptor. Cell. 1992 Aug 21;70(4):585–593. doi: 10.1016/0092-8674(92)90428-f. [DOI] [PubMed] [Google Scholar]
  36. Taniguchi T. Cytokine signaling through nonreceptor protein tyrosine kinases. Science. 1995 Apr 14;268(5208):251–255. doi: 10.1126/science.7716517. [DOI] [PubMed] [Google Scholar]
  37. Veillette A., Bookman M. A., Horak E. M., Bolen J. B. The CD4 and CD8 T cell surface antigens are associated with the internal membrane tyrosine-protein kinase p56lck. Cell. 1988 Oct 21;55(2):301–308. doi: 10.1016/0092-8674(88)90053-0. [DOI] [PubMed] [Google Scholar]
  38. Whitaker S., Geck P., Medveczky M. M., Cus J., Kung S. H., Lund T., Medveczky P. G. A polycistronic transcript in transformed cells encodes the dihydrofolate reductase of herpesvirus saimiri. Virus Genes. 1995;10(2):163–172. doi: 10.1007/BF01702597. [DOI] [PubMed] [Google Scholar]

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