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. 1983 Sep;47(3):487–494. doi: 10.1128/jvi.47.3.487-494.1983

Nonviable mutants of simian virus 40 with deletions near the 3' end of gene A define a function for large T antigen required after onset of viral DNA replication.

J Tornow, C N Cole
PMCID: PMC255290  PMID: 6312080

Abstract

Deletion mutants of simian virus 40 (SV40) with lesions at the three DdeI sites near the 3' end of the early region were constructed. Mutants with deletions at 0.203 and 0.219 map units (mu) which did not change the large T antigen reading frame were viable. This extends slightly the upstream boundary for the location of viable mutants with deletions in the 3' end of the A gene. Mutants with frameshift deletions at 0.193 and 0.219 mu were nonviable. These are the first nonviable mutants with deletions in this portion of the A gene. None of the three nonviable mutants with deletions at 0.219 mu produced progeny viral DNA. These three mutants all used the alternate reading frame located in this portion of the SV40 early region. The mutant with a deletion at 0.193 mu, dlA2459, was positive for viral DNA replication and was defective for adenovirus helper function. All of these mutations were located in the portion of the SV40 large T antigen which has no homology to the polyoma T antigens. These results indicate that this portion of large T antigen is required for some late step in the viral growth cycle and suggest that adenovirus helper function is required for productive infection by SV40.

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Selected References

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  1. Alwine J. C., Khoury G. Effect of a tsA mutation of simian virus 40 late gene expression: variations between host cell lines. J Virol. 1980 Feb;33(2):920–925. doi: 10.1128/jvi.33.2.920-925.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Alwine J. C., Khoury G. Simian virus 40-associated small RNA: mapping on the simian virus 40 genome and characterization of its synthesis. J Virol. 1980 Dec;36(3):701–708. doi: 10.1128/jvi.36.3.701-708.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Alwine J. C., Reed S. I., Stark G. R. Characterization of the autoregulation of simian virus 40 gene A. J Virol. 1977 Oct;24(1):22–27. doi: 10.1128/jvi.24.1.22-27.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bogenhagen D. F., Sakonju S., Brown D. D. A control region in the center of the 5S RNA gene directs specific initiation of transcription: II. The 3' border of the region. Cell. 1980 Jan;19(1):27–35. doi: 10.1016/0092-8674(80)90385-2. [DOI] [PubMed] [Google Scholar]
  6. Carbon J., Shenk T. E., Berg P. Biochemical procedure for production of small deletions in simian virus 40 DNA. Proc Natl Acad Sci U S A. 1975 Apr;72(4):1392–1396. doi: 10.1073/pnas.72.4.1392. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Chou J. Y., Martin R. G. Complementation analysis of simian virus 40 mutants. J Virol. 1974 May;13(5):1101–1109. doi: 10.1128/jvi.13.5.1101-1109.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Clewell D. B., Helinski D. R. Properties of a supercoiled deoxyribonucleic acid-protein relaxation complex and strand specificity of the relaxation event. Biochemistry. 1970 Oct 27;9(22):4428–4440. doi: 10.1021/bi00824a026. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. Cole C. N., Landers T., Goff S. P., Manteuil-Brutlag S., Berg P. Physical and genetic characterization of deletion mutants of simian virus 40 constructed in vitro. J Virol. 1977 Oct;24(1):277–294. doi: 10.1128/jvi.24.1.277-294.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Cosman D. J., Tevethia M. J. Characterization of a temperature-sensitive, DNA-positive, nontransforming mutant of simian virus 40. Virology. 1981 Jul 30;112(2):605–624. doi: 10.1016/0042-6822(81)90306-8. [DOI] [PubMed] [Google Scholar]
  12. Cowan K., Tegtmeyer P., Anthony D. D. Relationship of replication and transcription of Simian Virus 40 DNA. Proc Natl Acad Sci U S A. 1973 Jul;70(7):1927–1930. doi: 10.1073/pnas.70.7.1927. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Crawford L. V., Cole C. N., Smith A. E., Paucha E., Tegtmeyer P., Rundell K., Berg P. Organization and expression of early genes of simian virus 40. Proc Natl Acad Sci U S A. 1978 Jan;75(1):117–121. doi: 10.1073/pnas.75.1.117. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Denhardt D. T., Crawford L. V. Simian virus 40 T-antigen: identification of tryptic peptides in the C-terminal region and definition of the reading frame. J Virol. 1980 May;34(2):315–329. doi: 10.1128/jvi.34.2.315-329.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. 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]
  16. Fey G., Lewis J. B., Grodzicker T., Bothwell A. Characterization of a fused protein specified by the adenovirus type 2-simian virus 40 hybrid Ad2+ND1 dp2. J Virol. 1979 Apr;30(1):201–217. doi: 10.1128/jvi.30.1.201-217.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. 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]
  18. Friedmann T., Esty A., LaPorte P., Deininger P. The nucleotide sequence and genome organization of the polyoma early region: extensive nucleotide and amino acid homology with SV40. Cell. 1979 Jul;17(3):715–724. doi: 10.1016/0092-8674(79)90278-2. [DOI] [PubMed] [Google Scholar]
  19. Grodzicker T., Lewis J. B., Anderson C. W. Conditional lethal mutants of adenovirus type 2-simian virus 40 hybrids. II. Ad2+ND1 host-range mutants that synthesize fragments of the Ad2+ND1 30K protein. J Virol. 1976 Aug;19(2):559–571. doi: 10.1128/jvi.19.2.559-571.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hatanaka M., Dulbecco R. Induction of DNA synthesis by SV40. Proc Natl Acad Sci U S A. 1966 Aug;56(2):736–740. doi: 10.1073/pnas.56.2.736. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kimura G., Dulbecco R. A temperature-sensitive mutant of simian virus 40 affecting transforming ability. Virology. 1973 Apr;52(2):529–534. doi: 10.1016/0042-6822(73)90348-6. [DOI] [PubMed] [Google Scholar]
  22. Kimura G., Itagaki A. Initiation and maintenance of cell transformation by simian virus 40: a viral genetic property. Proc Natl Acad Sci U S A. 1975 Feb;72(2):673–677. doi: 10.1073/pnas.72.2.673. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Kit S. Viral-induced enzymes and the problem of viral oncogenesis. Adv Cancer Res. 1968;11:73–221. doi: 10.1016/s0065-230x(08)60388-7. [DOI] [PubMed] [Google Scholar]
  24. Klessig D. F., Anderson C. W. Block to multiplication of adenovirus serotype 2 in monkey cells. J Virol. 1975 Dec;16(6):1650–1668. doi: 10.1128/jvi.16.6.1650-1668.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Lai C. J., Nathans D. Deletion mutants of simian virus 40 generated by enzymatic excision of DNA segments from the viral genome. J Mol Biol. 1974 Oct 15;89(1):179–193. doi: 10.1016/0022-2836(74)90169-7. [DOI] [PubMed] [Google Scholar]
  26. Mandel M., Higa A. Calcium-dependent bacteriophage DNA infection. J Mol Biol. 1970 Oct 14;53(1):159–162. doi: 10.1016/0022-2836(70)90051-3. [DOI] [PubMed] [Google Scholar]
  27. Mark D. F., Berg P. A third splice site in SV40 early mRNA. Cold Spring Harb Symp Quant Biol. 1980;44(Pt 1):55–62. doi: 10.1101/sqb.1980.044.01.008. [DOI] [PubMed] [Google Scholar]
  28. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  29. Mertz J. E., Berg P. Defective simian virus 40 genomes: isolation and growth of individual clones. Virology. 1974 Nov;62(1):112–124. doi: 10.1016/0042-6822(74)90307-9. [DOI] [PubMed] [Google Scholar]
  30. Parker R. C., Watson R. M., Vinograd J. Mapping of closed circular DNAs by cleavage with restriction endonucleases and calibration by agarose gel electrophoresis. Proc Natl Acad Sci U S A. 1977 Mar;74(3):851–855. doi: 10.1073/pnas.74.3.851. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Pintel D., Bouck N., di Mayorca G. Separation of lytic and transforming functions of the simian virus 40 A region: two mutants which are temperature sensitive for lytic functions have opposite effects on transformation. J Virol. 1981 May;38(2):518–528. doi: 10.1128/jvi.38.2.518-528.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Pintel D., Bouck N., di Mayorca G., Thimmappaya B., Swerdlow B., Shenk T. SV40 mutant tsA1499 is heat-sensitive for lytic growth but generates cold-sensitive rat-cell transformants. Cold Spring Harb Symp Quant Biol. 1980;44(Pt 1):305–309. doi: 10.1101/sqb.1980.044.01.035. [DOI] [PubMed] [Google Scholar]
  33. Polvino-Bodnar M., Cole C. N. Construction and characterization of viable deletion mutants of simian virus 40 lacking sequences near the 3' end of the early region. J Virol. 1982 Aug;43(2):489–502. doi: 10.1128/jvi.43.2.489-502.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. RABSON A. S., O'CONOR G. T., BEREZESKY I. K., PAUL F. J. ENHANCEMENT OF ADENOVIRUS GROWTH IN AFRICAN GREEN MONKEY KIDNEY CELL CULTURES BY SV40. Proc Soc Exp Biol Med. 1964 May;116:187–190. doi: 10.3181/00379727-116-29197. [DOI] [PubMed] [Google Scholar]
  35. 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]
  36. Reed S. I., Stark G. R., Alwine J. C. Autoregulation of simian virus 40 gene A by T antigen. Proc Natl Acad Sci U S A. 1976 Sep;73(9):3083–3087. doi: 10.1073/pnas.73.9.3083. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Scheidtmann K. H., Echle B., Walter G. Simian virus 40 large T antigen is phosphorylated at multiple sites clustered in two separate regions. J Virol. 1982 Oct;44(1):116–133. doi: 10.1128/jvi.44.1.116-133.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Shortle D., Nathans D. Local mutagenesis: a method for generating viral mutants with base substitutions in preselected regions of the viral genome. Proc Natl Acad Sci U S A. 1978 May;75(5):2170–2174. doi: 10.1073/pnas.75.5.2170. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Soeda E., Arrand J. R., Smolar N., Walsh J. E., Griffin B. E. Coding potential and regulatory signals of the polyoma virus genome. Nature. 1980 Jan 31;283(5746):445–453. doi: 10.1038/283445a0. [DOI] [PubMed] [Google Scholar]
  40. Soprano K. J., Dev V. G., Croce C. M., Baserga R. Reactivation of silent rRNA genes by simian virus 40 in human-mouse hybrid cells. Proc Natl Acad Sci U S A. 1979 Aug;76(8):3885–3889. doi: 10.1073/pnas.76.8.3885. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  42. Takemoto K. K., Kirschstein R. L., Habel K. Mutants of simian virus 40 differing in plaque size, oncogenicity, and heat sensitivity. J Bacteriol. 1966 Oct;92(4):990–994. doi: 10.1128/jb.92.4.990-994.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Tegtmeyer P. Function of simian virus 40 gene A in transforming infection. J Virol. 1975 Mar;15(3):613–618. doi: 10.1128/jvi.15.3.613-618.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Tegtmeyer P., Schwartz M., Collins J. K., Rundell K. Regulation of tumor antigen synthesis by simain virus 40 gene A. J Virol. 1975 Jul;16(1):168–178. doi: 10.1128/jvi.16.1.168-178.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Tegtmeyer P. Simian virus 40 deoxyribonucleic acid synthesis: the viral replicon. J Virol. 1972 Oct;10(4):591–598. doi: 10.1128/jvi.10.4.591-598.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. 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]

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