Abstract
The phosphorylation sites of simian virus 40 (SV40) large tumor (T) antigens have been extensively studied in productive infection of monkey cells. In this study, we analyzed the phosphorylation sites of large T antigen from SV40-infected nonpermissive mouse cells by partial proteolysis fingerprints and analysis of the phosphoamino acids present in the resulting fragments. The wild-type virus and deletion mutants (dl1263, dl1265, dl2194, and dl2198) were used for infection. On the basis of our results and published data (M. Schwyzer, R. Weil, and H. Zuber, J. Biol. Chem. 225:5627-5634, 1980), a cleavage map of large T antigen was established. It was reported that at least four sites of phosphorylation were present. The amino-terminal part of the molecule contained both phosphoserine and phosphothreonine. One phosphothreonine residue was located in the prolinerich C-terminal end of the molecule at position 701 or 708. On the basis of the concensus as to the amino acid sequence surrounding the recognition sites for protein kinases, it was possible to more precisely locate this phosphothreonine at residue 701. Moreover, the C-terminal part of the molecule contained phosphoserine at a more internal position. In addition, this study firmly established the presence of a phosphothreonine in the N-terminal part of large T antigen. In conclusion, it was shown that the location of phosphorylation sites of large T antigen produced by nonpermissive mouse cells infected by SV40 is strikingly similar to that reported by other groups for large T antigen produced by SV40-infected permissive cells.
Full text
PDF










Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Bonner W. M., Stedman J. D. Efficient fluorography of 3H and 14C on thin layers. Anal Biochem. 1978 Aug 15;89(1):247–256. doi: 10.1016/0003-2697(78)90747-9. [DOI] [PubMed] [Google Scholar]
- Cole C. N., Crawford L. V., Berg P. Simian virus 40 mutants with deletions at the 3' end of the early region are defective in adenovirus helper function. J Virol. 1979 Jun;30(3):683–691. doi: 10.1128/jvi.30.3.683-691.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fanning E., Nowak B., Burger C. Detection and characterization of multiple forms of simian virus 40 large T antigen. J Virol. 1981 Jan;37(1):92–102. doi: 10.1128/jvi.37.1.92-102.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Feunteun J., Carmichael G., Nicolas J. C., Kress M. Mutant carrying deletions in the two simian virus 40 early genes. J Virol. 1981 Dec;40(3):625–634. doi: 10.1128/jvi.40.3.625-634.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fiers W., Contreras R., Haegemann G., Rogiers R., Van de Voorde A., Van Heuverswyn H., Van Herreweghe J., Volckaert G., Ysebaert M. Complete nucleotide sequence of SV40 DNA. Nature. 1978 May 11;273(5658):113–120. doi: 10.1038/273113a0. [DOI] [PubMed] [Google Scholar]
- Goldman N., Brown M., Khoury G. Modification of SV40 T antigen by poly ADP-ribosylation. Cell. 1981 May;24(2):567–572. doi: 10.1016/0092-8674(81)90347-0. [DOI] [PubMed] [Google Scholar]
- Greenspan D. S., Carroll R. B. Simian virus 40 large T antigen isoelectric focuses as multiple species with varying phosphate content. Virology. 1979 Dec;99(2):413–416. doi: 10.1016/0042-6822(79)90020-5. [DOI] [PubMed] [Google Scholar]
- Krebs E. G., Beavo J. A. Phosphorylation-dephosphorylation of enzymes. Annu Rev Biochem. 1979;48:923–959. doi: 10.1146/annurev.bi.48.070179.004423. [DOI] [PubMed] [Google Scholar]
- Kress M., May E., Cassingena R., May P. Simian virus 40-transformed cells express new species of proteins precipitable by anti-simian virus 40 tumor serum. J Virol. 1979 Aug;31(2):472–483. doi: 10.1128/jvi.31.2.472-483.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lai C. J., Goldman N. D., Khoury G. Functional similarity between the early antigens of simian virus 40 and human papovavirus BK. J Virol. 1979 Apr;30(1):141–147. doi: 10.1128/jvi.30.1.141-147.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Linke H. K., Hunter T., Walter G. Structural relationship between the 100,000- and 17,000- molecular-weight T antigens of simian virus 40 (SV40) as deduced by comparison with the SV40-specific proteins coded by the nondefective adenovirus type 2-SV40 hybrid viruses. J Virol. 1979 Jan;29(1):390–394. doi: 10.1128/jvi.29.1.390-394.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mann K., Hunter T. Phosphorylation of SV40 large T antigen in SV40 nucleoprotein complexes. Virology. 1980 Dec;107(2):526–532. doi: 10.1016/0042-6822(80)90320-7. [DOI] [PubMed] [Google Scholar]
- May E., Kress M., Daya-Grosjean L., Monier R., May P. Mapping of the viral mRNA encoding a super-T antigen of 115,000 daltons expressed in simian virus 40-transformed rat cell lines. J Virol. 1981 Jan;37(1):24–35. doi: 10.1128/jvi.37.1.24-35.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- May E., May P., Weil R. "Early" virus-specific RNA may contain information necessary for chromosome replication and mitosis induced by Simian Virus 40. Proc Natl Acad Sci U S A. 1973 Jun;70(6):1654–1658. doi: 10.1073/pnas.70.6.1654. [DOI] [PMC free article] [PubMed] [Google Scholar]
- McCormick F., Chaudry F., Harvey R., Smith R., Rigby P. W., Paucha E., Smith A. E. T antigens of SV40-transformed cells. Cold Spring Harb Symp Quant Biol. 1980;44(Pt 1):171–178. doi: 10.1101/sqb.1980.044.01.020. [DOI] [PubMed] [Google Scholar]
- Mercier J. C. Phosphorylation of caseins, present evidence for an amino acid triplet code posttranslationally recognized by specific kinases. Biochimie. 1981 Jan;63(1):1–17. doi: 10.1016/s0300-9084(81)80141-1. [DOI] [PubMed] [Google Scholar]
- Montenarh M., Henning R. Simian virus 40 T-antigen phosphorylation is variable. FEBS Lett. 1980 May 19;114(1):107–110. doi: 10.1016/0014-5793(80)80870-2. [DOI] [PubMed] [Google Scholar]
- Paucha E., Mellor A., Harvey R., Smith A. E., Hewick R. M., Waterfield M. D. Large and small tumor antigens from simian virus 40 have identical amino termini mapping at 0.65 map units. Proc Natl Acad Sci U S A. 1978 May;75(5):2165–2169. doi: 10.1073/pnas.75.5.2165. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Reddy V. B., Thimmappaya B., Dhar R., Subramanian K. N., Zain B. S., Pan J., Ghosh P. K., Celma M. L., Weissman S. M. The genome of simian virus 40. Science. 1978 May 5;200(4341):494–502. doi: 10.1126/science.205947. [DOI] [PubMed] [Google Scholar]
- Rundell K., Collins J. K., Tegtmeyer P., Ozer H. L., Lai C. J., Nathans D. Identification of simian virus 40 protein A. J Virol. 1977 Feb;21(2):636–646. doi: 10.1128/jvi.21.2.636-646.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Scheidtmann K. H., Kaiser A., Carbone A., Walter G. Phosphorylation of threonine in the proline-rich carboxy-terminal region of simian virus 40 large T antigen. J Virol. 1981 Apr;38(1):59–69. doi: 10.1128/jvi.38.1.59-69.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schwyzer M., Weil R., Frank G., Zuber H. Amino acid sequence analysis of fragments generated by partial proteolysis from large simian virus 40 tumor antigen. J Biol Chem. 1980 Jun 25;255(12):5627–5634. [PubMed] [Google Scholar]
- Seif I., Khoury G., Dhar R. The genome of human papovavirus BKV. Cell. 1979 Dec;18(4):963–977. doi: 10.1016/0092-8674(79)90209-5. [DOI] [PubMed] [Google Scholar]
- Small D., Chou P. Y., Fasman G. D. Occurrence of phosphorylated residues in predicted beta-turns: implications for beta-turn participation in control mechanisms. Biochem Biophys Res Commun. 1977 Nov 7;79(1):341–346. doi: 10.1016/0006-291x(77)90101-2. [DOI] [PubMed] [Google Scholar]
- Smith A. E., Smith R., Paucha E. Extraction and fingerprint analysis of simian virus 40 large and small T-antigens. J Virol. 1978 Oct;28(1):140–153. doi: 10.1128/jvi.28.1.140-153.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tegtmeyer P., Rundell K., Collins J. K. Modification of simian virus 40 protein A. J Virol. 1977 Feb;21(2):647–657. doi: 10.1128/jvi.21.2.647-657.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tournier P., Cassingena R., Wicker R., Coppey J., Suarez H. Etude du mécanisme de l'induction chez des cellules de hamster syrien transformées par le virus SV40. I. Propriétés d'une lignée cellulaire clonale. Int J Cancer. 1967 Mar 15;2(2):117–132. doi: 10.1002/ijc.2910020207. [DOI] [PubMed] [Google Scholar]
- Van Heuverswyn H., Cole C., Berg P., Fiers W. Nucleotide sequence analysis of two simian virus 40 mutants with deletions in the region coding for the carboxyl terminus of the T antigen. J Virol. 1979 Jun;30(3):936–941. doi: 10.1128/jvi.30.3.936-941.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Van Heuverswyn H., Fiers W. Nucleotide sequence of the Hind-C fragment of simian virus 40 DNA. Comparison of the 5'-untranslated region of wild-type virus and of some deletion Mutants. Eur J Biochem. 1979 Oct;100(1):51–60. doi: 10.1111/j.1432-1033.1979.tb02032.x. [DOI] [PubMed] [Google Scholar]
- Van Roy F., Fransen L., Fiers W. Phosphorylation patterns of tumour antigens in cells lytically infected or transformed by simian virus 40. J Virol. 1981 Oct;40(1):28–44. doi: 10.1128/jvi.40.1.28-44.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Walter G., Flory P. J., Jr Phosphorylation of SV40 large T antigen. Cold Spring Harb Symp Quant Biol. 1980;44(Pt 1):165–169. doi: 10.1101/sqb.1980.044.01.019. [DOI] [PubMed] [Google Scholar]






