Abstract
The bulk of simian virus 40 (SV40) large T antigen in SV40-infected and -transformed cells localizes within the cell nucleus, while a minor fraction specifically associates with the plasma membrane (PM) and is exposed on the cell surface. PM-associated large T seems to span the lipid bilayer but, on the other hand, does not display typical features of a transmembrane protein. To further characterize the postulated transmembrane orientation of large T, we asked whether all large T molecules associated with the plasma membrane indeed are exposed on the cell surface. We compared the amount of cell surface-exposed large T, determined on living cells by a sensitive 3H-protein A-binding assay and by external immunoprecipitation, with that of total PM-associated large T extracted from isolated PM. We demonstrate that in mKSA cells (SV40-transformed BALB/c mouse fibroblasts), total PM-associated large T accounted for a substantial portion (ca. 2%) of total cellular large T. However, only 0.1 to 0.2% of it could be detected on the cell surface. Thus, only a minor fraction of PM-associated large T (less than 10%) is exposed on the surface of these cells. Interior PM-associated large T is stably associated with the plasma membrane, while the small fraction of surface-exposed large T is rapidly released from the cell surface.
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- Ben-Ze'ev A., Duerr A., Solomon F., Penman S. The outer boundary of the cytoskeleton: a lamina derived from plasma membrane proteins. Cell. 1979 Aug;17(4):859–865. doi: 10.1016/0092-8674(79)90326-x. [DOI] [PubMed] [Google Scholar]
- Bishop J. M. Viral oncogenes. Cell. 1985 Aug;42(1):23–38. doi: 10.1016/s0092-8674(85)80098-2. [DOI] [PubMed] [Google Scholar]
- Butel J. S., Jarvis D. L. The plasma-membrane-associated form of SV40 large tumor antigen: biochemical and biological properties. Biochim Biophys Acta. 1986 Oct 28;865(2):171–195. doi: 10.1016/0304-419x(86)90027-2. [DOI] [PubMed] [Google Scholar]
- Carter W. G., Hakomori S. A new cell surface, detergent-insoluble glycoprotein matrix of human and hamster fibroblasts. The role of disulfide bonds in stabilization of the matrix. J Biol Chem. 1981 Jul 10;256(13):6953–6960. [PubMed] [Google Scholar]
- DeLeo A. B., Shiku H., Takahashi T., John M., Old L. J. Cell surface antigens of chemically induced sarcomas of the mouse. I. Murine leukemia virus-related antigens and alloantigens on cultured fibroblasts and sarcoma cells: description of a unique antigen on BALB/c Meth A sarcoma. J Exp Med. 1977 Sep 1;146(3):720–734. doi: 10.1084/jem.146.3.720. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Deppert W., Hanke K., Henning R. Simian virus 40 T-antigen-related cell surface antigen: serological demonstration on simian virus 40-transformed monolayer cells in situ. J Virol. 1980 Aug;35(2):505–518. doi: 10.1128/jvi.35.2.505-518.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Deppert W., Walser A., Klockmann U. A subclass of the adenovirus 72K DNA binding protein specifically associating with the cytoskeletal framework of the plasma membrane. Virology. 1988 Aug;165(2):457–468. doi: 10.1016/0042-6822(88)90589-2. [DOI] [PubMed] [Google Scholar]
- Deppert W., Walter G. Domains of simian virus 40 large T-antigen exposed on the cell surface. Virology. 1982 Oct 15;122(1):56–70. doi: 10.1016/0042-6822(82)90377-4. [DOI] [PubMed] [Google Scholar]
- Flaherty L., Zimmerman D. Surface mapping of mouse thymocytes. Proc Natl Acad Sci U S A. 1979 Apr;76(4):1990–1993. doi: 10.1073/pnas.76.4.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Graham F. L., Smiley J., Russell W. C., Nairn R. Characteristics of a human cell line transformed by DNA from human adenovirus type 5. J Gen Virol. 1977 Jul;36(1):59–74. doi: 10.1099/0022-1317-36-1-59. [DOI] [PubMed] [Google Scholar]
- Gurney E. G., Tamowski S., Deppert W. Antigenic binding sites of monoclonal antibodies specific for simian virus 40 large T antigen. J Virol. 1986 Mar;57(3):1168–1172. doi: 10.1128/jvi.57.3.1168-1172.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jarvis D. L., Chan W. K., Estes M. K., Butel J. S. The cellular secretory pathway is not utilized for biosynthesis, modification, or intracellular transport of the simian virus 40 large tumor antigen. J Virol. 1987 Dec;61(12):3950–3959. doi: 10.1128/jvi.61.12.3950-3959.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kit S., Kurimura T., Dubbs D. R. Transplantable mouse tumor line induced by injection of SV40-transformed mouse kidney cells. Int J Cancer. 1969 Jul 15;4(4):384–392. doi: 10.1002/ijc.2910040403. [DOI] [PubMed] [Google Scholar]
- Klockmann U., Deppert W. Acylated simian virus 40 large T-antigen: a new subclass associated with a detergent-resistant lamina of the plasma membrane. EMBO J. 1983;2(7):1151–1157. doi: 10.1002/j.1460-2075.1983.tb01560.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Klockmann U., Deppert W. Evidence for transmembrane orientation of acylated simian virus 40 large T antigen. J Virol. 1985 Nov;56(2):541–548. doi: 10.1128/jvi.56.2.541-548.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lanford R. E., Butel J. S. Antigenic relationship of SV40 early proteins to purified large T polypeptide. Virology. 1979 Sep;97(2):295–306. doi: 10.1016/0042-6822(79)90341-6. [DOI] [PubMed] [Google Scholar]
- MacArthur H., Walter G. Monoclonal antibodies specific for the carboxy terminus of simian virus 40 large T antigen. J Virol. 1984 Nov;52(2):483–491. doi: 10.1128/jvi.52.2.483-491.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mescher M. F., Jose M. J., Balk S. P. Actin-containing matrix associated with the plasma membrane of murine tumour and lymphoid cells. Nature. 1981 Jan 15;289(5794):139–144. doi: 10.1038/289139a0. [DOI] [PubMed] [Google Scholar]
- Rinke Y., Deppert W. Quantitative analysis of cell surface-associated SV40 large T antigen using a newly developed 3H-protein A binding assay. Virology. 1989 Jun;170(2):424–432. doi: 10.1016/0042-6822(89)90433-9. [DOI] [PubMed] [Google Scholar]
- Santos M., Butel J. S. Antigenic structure of simian virus 40 large tumor antigen and association with cellular protein p53 on the surfaces of simian virus 40-infected and -transformed cells. J Virol. 1984 Aug;51(2):376–383. doi: 10.1128/jvi.51.2.376-383.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Santos M., Butel J. S. Association of SV40 large tumor antigen and cellular proteins on the surface of SV40-transformed mouse cells. Virology. 1982 Jul 15;120(1):1–17. doi: 10.1016/0042-6822(82)90002-2. [DOI] [PubMed] [Google Scholar]
- Santos M., Butel J. S. Dynamic nature of the association of large tumor antigen and p53 cellular protein with the surfaces of simian virus 40-transformed cells. J Virol. 1984 Jan;49(1):50–56. doi: 10.1128/jvi.49.1.50-56.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Santos M., Butel J. S. Surface T-antigen expression in simian virus 40-transformed mouse cells: correlation with cell growth rate. Mol Cell Biol. 1985 May;5(5):1051–1057. doi: 10.1128/mcb.5.5.1051. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sharma S., Rodgers L., Brandsma J., Gething M. J., Sambrook J. SV40 T antigen and the exocytotic pathway. EMBO J. 1985 Jun;4(6):1479–1489. doi: 10.1002/j.1460-2075.1985.tb03806.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Soule H. R., Butel J. S. Subcellular Localization of simian virus 40 large tumor antigen. J Virol. 1979 May;30(2):523–532. doi: 10.1128/jvi.30.2.523-532.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Stahl H., Knippers R. The simian virus 40 large tumor antigen. Biochim Biophys Acta. 1987 Oct 9;910(1):1–10. doi: 10.1016/0167-4781(87)90088-1. [DOI] [PubMed] [Google Scholar]
- Tevethia M. J., Anderson R. W., Tevethia S. S., Simmons D., Feunteun J., Cole C. Influence of amino acids encoded in the 3' open reading frame of the SV40 early region on transformation and antigenicity of large T antigen. Virology. 1986 Apr 30;150(2):361–372. doi: 10.1016/0042-6822(86)90301-6. [DOI] [PubMed] [Google Scholar]
- Whittaker L., Fuks A., Hand R. Plasma membrane orientation of simian virus 40 T antigen in three transformed cell lines mapped with monoclonal antibodies. J Virol. 1985 Feb;53(2):366–373. doi: 10.1128/jvi.53.2.366-373.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]




