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. 1979 Apr;30(1):279–296. doi: 10.1128/jvi.30.1.279-296.1979

Simian virus 40 early mRNA's. I. Genomic localization of 3' and 5' termini and two major splices in mRNA from transformed and lytically infected cells.

V B Reddy, P K Ghosh, P Lebowitz, M Piatak, S M Weissman
PMCID: PMC353322  PMID: 90157

Abstract

We have studied the structure of polyadenylated virus-specific cytoplasmic mRNA's in mouse and human cells transformed by simian virus 40 and in monkey cells infected with simian virus 40 in the presence of cytosine arabinoside by means of reverse transcriptase-catalyzed complementary DNA synthesis and complementary DNA sequencing. Abundant mRNA species containing splices from residues 4490 to 4557 (0.533 to 0.546 map units [m.u.]) and 4490 to 4837 (0.533 to 0.600 m.u.) were identified in both transformed and infected cells. Two principal reverse transcriptase stops were observed at the 5' termini of these mRNA's, both occurring with approximately equal frequency. The most distal of these stops was localized at residues 5152 to 5154 (0.660 m.u.), and the second was at residues 5147 to 5148 (0.659 m.u.). Several additional minor stops, between approximately 0.62 and 0.65 m.u., were also found on complementary DNA copied from transformed cell mRNA; in contrast, only one additional stop was present on complementary DNA copied from early lytic mRNA. These data suggest the presence of a prinicipal 5' terminus of early lytic and transformed cell mRNA's at residues 5152 to 5154 and raise the possibility of additional 5' termini at one or more locations in the 0.62 to 0.659 m.u. region of these mRNA's. Transformed cell mRNA was also found to contain a single 3' terminus at positions 2504 and 2505 (0.153 m.u.); termini lying beyond this site were not detected.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Abrahams P. J., Mulder C., Van De Voorde A., Warnaar S. O., van der Eb A. J. Transformation of primary rat kidney cells by fragments of simian virus 40 DNA. J Virol. 1975 Oct;16(4):818–823. doi: 10.1128/jvi.16.4.818-823.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Abrahams P. J., Van der Eb A. J. In vitro transformation of rat and mouse cells by DNA from simian virus 40. J Virol. 1975 Jul;16(1):206–209. doi: 10.1128/jvi.16.1.206-209.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Baralle F. E., Brownlee G. G. AUG is the only recognisable signal sequence in the 5' non-coding regions of eukaryotic mRNA. Nature. 1978 Jul 6;274(5666):84–87. doi: 10.1038/274084a0. [DOI] [PubMed] [Google Scholar]
  4. Berk A. J., Sharp P. A. Spliced early mRNAs of simian virus 40. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1274–1278. doi: 10.1073/pnas.75.3.1274. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Blackburn P., Wilson G., Moore S. Ribonuclease inhibitor from human placenta. Purification and properties. J Biol Chem. 1977 Aug 25;252(16):5904–5910. [PubMed] [Google Scholar]
  6. Blakesley R. W., Wells R. D. 'Single-stranded' DNA from phiX174 and M13 is cleaved by certain restriction endonucleases. Nature. 1975 Oct 2;257(5525):421–422. doi: 10.1038/257421a0. [DOI] [PubMed] [Google Scholar]
  7. Botchan M., Topp W., Sambrook J. The arrangement of simian virus 40 sequences in the DNA of transformed cells. Cell. 1976 Oct;9(2):269–287. doi: 10.1016/0092-8674(76)90118-5. [DOI] [PubMed] [Google Scholar]
  8. Brack C., Tonegawa S. Variable and constant parts of the immunoglobulin light chain gene of a mouse myeloma cell are 1250 nontranslated bases apart. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5652–5656. doi: 10.1073/pnas.74.12.5652. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Breathnach R., Benoist C., O'Hare K., Gannon F., Chambon P. Ovalbumin gene: evidence for a leader sequence in mRNA and DNA sequences at the exon-intron boundaries. Proc Natl Acad Sci U S A. 1978 Oct;75(10):4853–4857. doi: 10.1073/pnas.75.10.4853. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Breathnach R., Mandel J. L., Chambon P. Ovalbumin gene is split in chicken DNA. Nature. 1977 Nov 24;270(5635):314–319. doi: 10.1038/270314a0. [DOI] [PubMed] [Google Scholar]
  11. Brugge J. S., Butel J. S. Role of simian virus 40 gene A function in maintenance of transformation. J Virol. 1975 Mar;15(3):619–635. doi: 10.1128/jvi.15.3.619-635.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Chou J. Y., Martin R. G. Products of complementation between temperature-sensitive mutants of simian virus 40. J Virol. 1975 Jan;15(1):127–136. doi: 10.1128/jvi.15.1.127-136.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. 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]
  14. 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]
  15. Dhar R., Subramanian K. N., Pan J., Weissman S. M. Nucleotide sequence of a fragment of SV40 DNA that contains the origin of DNA replication and specifies the 5' ends of "early" and "late" viral RNA. IV. Localization of the SV40 DNA complementary to the 5' ends of viral mRNA. J Biol Chem. 1977 Jan 10;252(1):368–376. [PubMed] [Google Scholar]
  16. Dhar R., Subramanian K., Zain B. S., Pan J., Weissman S. M. Nucleotide sequence about the 3' terminus of SV40 DNA transcripts and the region where DNA synthesis is initiated. Cold Spring Harb Symp Quant Biol. 1975;39(Pt 1):153–160. doi: 10.1101/sqb.1974.039.01.021. [DOI] [PubMed] [Google Scholar]
  17. Dickson R. C., Abelson J., Barnes W. M., Reznikoff W. S. Genetic regulation: the Lac control region. Science. 1975 Jan 10;187(4171):27–35. doi: 10.1126/science.1088926. [DOI] [PubMed] [Google Scholar]
  18. Efstratiadis A., Kafatos F. C., Maniatis T. The primary structure of rabbit beta-globin mRNA as determined from cloned DNA. Cell. 1977 Apr;10(4):571–585. doi: 10.1016/0092-8674(77)90090-3. [DOI] [PubMed] [Google Scholar]
  19. Ferdinand F. J., Brown M., Khoury G. Characterization of early simian virus 40 transcriptional complexes: late transcription in the absence of detectable DNA replication. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5443–5447. doi: 10.1073/pnas.74.12.5443. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. 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]
  21. Ghosh P. K., Reddy V. B., Swinscoe J., Choudary P. V., Lebowitz P., Weissman S. M. The 5'-terminal leader sequence of late 16 S mRNA from cells infected with simian virus 40. J Biol Chem. 1978 May 25;253(10):3643–3647. [PubMed] [Google Scholar]
  22. Horiuchi K., Zinder N. D. Site-specific cleavage of single-stranded DNA by a Hemophilus restriction endonuclease. Proc Natl Acad Sci U S A. 1975 Jul;72(7):2555–2558. doi: 10.1073/pnas.72.7.2555. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Khoury G., Byrne J. C., Martin M. A. Patterns of Simian Virus 40 DNA transcription after acute infection of permissive and nonpermissive cells. Proc Natl Acad Sci U S A. 1972 Jul;69(7):1925–1928. doi: 10.1073/pnas.69.7.1925. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Khoury G., Howley P., Nathans D., Martin M. Posttranscriptional selection of simian virus 40-specific RNA. J Virol. 1975 Feb;15(2):433–437. doi: 10.1128/jvi.15.2.433-437.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Khoury G., Martin M. A., Lee T. N., Danna K. J., Nathans D. A map of simian virus 40 transcription sites expressed in productively infected cells. J Mol Biol. 1973 Aug 5;78(2):377–389. doi: 10.1016/0022-2836(73)90123-x. [DOI] [PubMed] [Google Scholar]
  26. Khoury G., May E. Regulation of early and late simian virus 40 transcription: overproduction of early viral RNA in the absence of a functional T-antigen. J Virol. 1977 Jul;23(1):167–176. doi: 10.1128/jvi.23.1.167-176.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Kim C. H., Sarma R. H. Spatial configuration of mRNA 5'-terminus. Nature. 1977 Nov 17;270(5634):223–227. doi: 10.1038/270223a0. [DOI] [PubMed] [Google Scholar]
  28. Lee T. N., Nathans D. A transcriptional map of the SV40 genome in transformed cell lines. Virology. 1975 Jan;63(1):263–272. doi: 10.1016/0042-6822(75)90390-6. [DOI] [PubMed] [Google Scholar]
  29. Martin R. G., Chou J. Y. Simian virus 40 functions required for the establishment and maintenance of malignant transformation. J Virol. 1975 Mar;15(3):599–612. doi: 10.1128/jvi.15.3.599-612.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Maxam A. M., Gilbert W. A new method for sequencing DNA. Proc Natl Acad Sci U S A. 1977 Feb;74(2):560–564. doi: 10.1073/pnas.74.2.560. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Mertz J. E., Berg P. Viable deletion mutants of simian virus 40: selective isolation by means of a restriction endonuclease from Hemophilus parainfluenzae. Proc Natl Acad Sci U S A. 1974 Dec;71(12):4879–4883. doi: 10.1073/pnas.71.12.4879. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Osborn M., Weber K. Simian virus 40 gene A function and maintenance of transformation. J Virol. 1975 Mar;15(3):636–644. doi: 10.1128/jvi.15.3.636-644.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Paucha E., Harvey R., Smith A. E. Cell-free synthesis of simian virus 40 T-antigens. J Virol. 1978 Oct;28(1):154–170. doi: 10.1128/jvi.28.1.154-170.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. 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]
  35. Peacock A. C., Dingman C. W. Molecular weight estimation and separation of ribonucleic acid by electrophoresis in agarose-acrylamide composite gels. Biochemistry. 1968 Feb;7(2):668–674. doi: 10.1021/bi00842a023. [DOI] [PubMed] [Google Scholar]
  36. Penman S. RNA metabolism in the HeLa cell nucleus. J Mol Biol. 1966 May;17(1):117–130. doi: 10.1016/s0022-2836(66)80098-0. [DOI] [PubMed] [Google Scholar]
  37. Prives C., Gilboa E., Revel M., Winocour E. Cell-free translation of simian virus 40 early messenger RNA coding for viral T-antigen. Proc Natl Acad Sci U S A. 1977 Feb;74(2):457–461. doi: 10.1073/pnas.74.2.457. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Proudfoot N. J. Complete 3' noncoding region sequences of rabbit and human beta-globin messenger RNAs. Cell. 1977 Apr;10(4):559–570. doi: 10.1016/0092-8674(77)90089-7. [DOI] [PubMed] [Google Scholar]
  39. Pyrrhic victories for the friends of whales. Nature. 1972 Jul 7;238(5358):4–4. doi: 10.1038/238004a0. [DOI] [PubMed] [Google Scholar]
  40. Rassoulzadegan M., Perbal B., Cuzin F. Growth control in simian virus 40-transformed rat cells: temperature-independent expression of the transformed phenotype in tsA transformants derived by agar selection. J Virol. 1978 Oct;28(1):1–5. doi: 10.1128/jvi.28.1.1-5.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. 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]
  42. 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]
  43. 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]
  44. Sambrook J., Sharp P. A., Keller W. Transcription of Simian virus 40. I. Separation of the strands of SV40 DNA and hybridization of the separated strands to RNA extracted from lytically infected and transformed cells. J Mol Biol. 1972 Sep 14;70(1):57–71. doi: 10.1016/0022-2836(72)90163-5. [DOI] [PubMed] [Google Scholar]
  45. Sambrook J., Sugden B., Keller W., Sharp P. A. Transcription of simian virus 40. 3. Mapping of "early" and "late" species of RNA. Proc Natl Acad Sci U S A. 1973 Dec;70(12):3711–3715. doi: 10.1073/pnas.70.12.3711. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Shenk T. E., Carbon J., Berg P. Construction and analysis of viable deletion mutants of simian virus 40. J Virol. 1976 May;18(2):664–671. doi: 10.1128/jvi.18.2.664-671.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Simmons D. T., Martin M. A. Common methionine-tryptic peptides near the amino-terminal end of primate papovavirus tumor antigens. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1131–1135. doi: 10.1073/pnas.75.3.1131. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Sleigh M. J., Topp W. C., Hanich R., Sambrook J. F. Mutants of SV40 with an altered small t protein are reduced in their ability to transform cells. Cell. 1978 May;14(1):79–88. doi: 10.1016/0092-8674(78)90303-3. [DOI] [PubMed] [Google Scholar]
  49. 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]
  50. 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]
  51. 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]
  52. 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]
  53. Tilghman S. M., Tiemeier D. C., Seidman J. G., Peterlin B. M., Sullivan M., Maizel J. V., Leder P. Intervening sequence of DNA identified in the structural portion of a mouse beta-globin gene. Proc Natl Acad Sci U S A. 1978 Feb;75(2):725–729. doi: 10.1073/pnas.75.2.725. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Tjian R. The binding site on SV40 DNA for a T antigen-related protein. Cell. 1978 Jan;13(1):165–179. doi: 10.1016/0092-8674(78)90147-2. [DOI] [PubMed] [Google Scholar]
  55. Tonegawa S., Maxam A. M., Tizard R., Bernard O., Gilbert W. Sequence of a mouse germ-line gene for a variable region of an immunoglobulin light chain. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1485–1489. doi: 10.1073/pnas.75.3.1485. [DOI] [PMC free article] [PubMed] [Google Scholar]
  56. Wiegand R. C., Godson G. N., Radding C. M. Specificity of the S1 nuclease from Aspergillus oryzae. J Biol Chem. 1975 Nov 25;250(22):8848–8855. [PubMed] [Google Scholar]
  57. Wilson J. T., Wilson L. B., deRiel J. K., Villa-komaroff L., Efstratiadis A., Forget B. G., Weissman S. M. Insertion of synthetic copies of human globin genes into bacterial plasmids. Nucleic Acids Res. 1978 Feb;5(2):563–581. doi: 10.1093/nar/5.2.563. [DOI] [PMC free article] [PubMed] [Google Scholar]
  58. Wilson J. T., deRiel J. K., Forget B. G., Marotta C. A., Weissman S. M. Nucleotide sequence of 3' untranslated portion of human alpha globin mRNA. Nucleic Acids Res. 1977 Jul;4(7):2353–2368. doi: 10.1093/nar/4.7.2353. [DOI] [PMC free article] [PubMed] [Google Scholar]
  59. Zain B. S., Dhar R., Weissman S. M., Lebowitz P., Lewis A. M., Jr Preferred site for initiation of RNA transcription by Escherichia coli RNA polymerase within the simian virus 40 DNA segment of the nondefective adenovirus-simian virus 40 hybrid viruses Ad2 + ND 1 and Ad2 + ND 3 . J Virol. 1973 May;11(5):682–693. doi: 10.1128/jvi.11.5.682-693.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]

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