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. 1979 Jul;76(7):3078–3082. doi: 10.1073/pnas.76.7.3078

Sequence heterogeneity at the 5' termini of late simian virus 40 19S and 16S mRNAs.

D Canaani, C Kahana, A Mukamel, Y Groner
PMCID: PMC383766  PMID: 226954

Abstract

The 5'-cap-containing leader sequence of the most abundant 19S and 16S mRNAs of simian virus 40 (SV40) was previously mapped between 0.67 and 0.76 map units. We now find that the two late mRNA species contain multiple 5' ends. Eight different RNase T2-resistant cap structures were identified:m7GpppmAmpU (47%); m7GpppmAmpUmpU (19%); m7GpppmAmpC (16%); m7GpppmAmpCmpA (5%); m7GpppmAmpG (6%); m7GpppGmpC (3%); m7GpppmAmGmpA (2%); m7GpppGmpCmpG (2%). Capped T1 oligonucleotides of 19S and 16S mRNAs have been isolated by two different procedures: (i) chromatography on a DEAE-cellulose column followed by paper electrophoresis and (ii) two-dimensional electrophoresis/homochromatography. Cap structures of the isolated 5' oligonucleotides were identified. Each of the major caps was found to be associated with a few differential 5' oligonucleotides, implying a vast heterogeneity at the termini of SV40 late mRNAs. The results suggest that on SV40 DNA, RNA polymerase II has a reportoire of initiation points. In most of the cases, initiation takes place with adenosine triphosphate followed by a pyrimidine. Alternatively, transcription may start at one specific point but a unique mechanism of processing generates heterogeneous populations of termini with a common 5' adenosine triphosphate.

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Selected References

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

  1. Aloni Y. Biogenesis and characterization of SV40 and polyoma RNAs in productively infected cells. Cold Spring Harb Symp Quant Biol. 1975;39(Pt 1):165–178. doi: 10.1101/sqb.1974.039.01.023. [DOI] [PubMed] [Google Scholar]
  2. Aloni Y., Dhar R., Laub O., Horowitz M., Khoury G. Novel mechanism for RNA maturation: the leader sequences of simian virus 40 mRNA are not transcribed adjacent to the coding sequences. Proc Natl Acad Sci U S A. 1977 Sep;74(9):3686–3690. doi: 10.1073/pnas.74.9.3686. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Berget S. M., Moore C., Sharp P. A. Spliced segments at the 5' terminus of adenovirus 2 late mRNA. Proc Natl Acad Sci U S A. 1977 Aug;74(8):3171–3175. doi: 10.1073/pnas.74.8.3171. [DOI] [PMC free article] [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. Bina-Stein M., Thoren M., Salzman N., Thomspon J. A. Rapid sequence determination of late simian virus 40 16S mRNA leader by using inhibitors of reverse transcriptase. Proc Natl Acad Sci U S A. 1979 Feb;76(2):731–735. doi: 10.1073/pnas.76.2.731. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bratosin S., Horowitz M., Laub O., Aloni Y. Electron microscopic evidence for splicing of SV40 late mRNAs. Cell. 1978 Apr;13(4):783–790. doi: 10.1016/0092-8674(78)90228-3. [DOI] [PubMed] [Google Scholar]
  7. Chow L. T., Broker T. R. The spliced structures of adenovirus 2 fiber message and the other late mRNAs. Cell. 1978 Oct;15(2):497–510. doi: 10.1016/0092-8674(78)90019-3. [DOI] [PubMed] [Google Scholar]
  8. Chow L. T., Gelinas R. E., Broker T. R., Roberts R. J. An amazing sequence arrangement at the 5' ends of adenovirus 2 messenger RNA. Cell. 1977 Sep;12(1):1–8. doi: 10.1016/0092-8674(77)90180-5. [DOI] [PubMed] [Google Scholar]
  9. Dhar R., Subramanian K. N., Pan J., Weissman S. M. Structure of a large segment of the genome of simian virus 40 that does not encode known proteins. Proc Natl Acad Sci U S A. 1977 Mar;74(3):827–831. doi: 10.1073/pnas.74.3.827. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Flavell A. J., Cowie A., Legon S., Kamen R. Multiple 5' terminal cap structures in late polyoma virus RNA. Cell. 1979 Feb;16(2):357–371. doi: 10.1016/0092-8674(79)90012-6. [DOI] [PubMed] [Google Scholar]
  11. Furuichi Y. "Pretranscriptional capping" in the biosynthesis of cytoplasmic polyhedrosis virus mRNA. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1086–1090. doi: 10.1073/pnas.75.3.1086. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Gelinas R. E., Roberts R. J. One predominant 5'-undecanucleotide in adenovirus 2 late messenger RNAs. Cell. 1977 Jul;11(3):533–544. doi: 10.1016/0092-8674(77)90071-x. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. Ghosh P. K., Reddy V. B., Swinscoe J., Lebowitz P., Weissman S. M. Heterogeneity and 5'-terminal structures of the late RNAs of simian virus 40. J Mol Biol. 1978 Dec 25;126(4):813–846. doi: 10.1016/0022-2836(78)90022-0. [DOI] [PubMed] [Google Scholar]
  15. Gilboa E., Prives C. L., Aviv H. Purification of SV-40 messenger RNA by hybridization to SV-40 DNA covalently bound to Sepharose. Biochemistry. 1975 Sep 23;14(19):4215–4220. doi: 10.1021/bi00690a010. [DOI] [PubMed] [Google Scholar]
  16. Gilham P. T., Rosenberg M. The isolation of 3'-terminal polynucleotides from RNA molecules. Biochim Biophys Acta. 1971 Aug 26;246(2):337–340. doi: 10.1016/0005-2787(71)90143-2. [DOI] [PubMed] [Google Scholar]
  17. Groner Y., Carmi P., Aloni Y. Capping structures of simian virus 40 19S and 16S mRNAs. Nucleic Acids Res. 1977 Nov;4(11):3959–3968. doi: 10.1093/nar/4.11.3959. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Groner Y., Gilboa E., Aviv H. Methylation and capping of RNA polymerase II primary transcripts by HeLa nuclear homogenates. Biochemistry. 1978 Mar 21;17(6):977–982. doi: 10.1021/bi00599a005. [DOI] [PubMed] [Google Scholar]
  19. Groner Y., Grosfeld H., Littauer U. Z. 5'-Capping structures of Artemia salina mRNA and the translational inhibition by cap analogs. Eur J Biochem. 1976 Dec;71(1):281–293. doi: 10.1111/j.1432-1033.1976.tb11114.x. [DOI] [PubMed] [Google Scholar]
  20. Groner Y., Hurwitz J. Synthesis of RNA containing a methylated blocked 5' terminus by HeLa nuclear homogenates. Proc Natl Acad Sci U S A. 1975 Aug;72(8):2930–2934. doi: 10.1073/pnas.72.8.2930. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Haegeman G., Fiers W. Characterization of the 5'-terminal capped structures of late simian virus 40-specific mRNA. J Virol. 1978 Mar;25(3):824–830. doi: 10.1128/jvi.25.3.824-830.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Haegeman G., Fiers W. Localization of the 5' terminus of late SV40 mRNA. Nucleic Acids Res. 1978 Jul;5(7):2359–2371. doi: 10.1093/nar/5.7.2359. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Hsu M. T., Ford J. Sequence arrangement of the 5' ends of simian virus 40 16S and 19S mRNAs. Proc Natl Acad Sci U S A. 1977 Nov;74(11):4982–4985. doi: 10.1073/pnas.74.11.4982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Klessig D. F. Two adenovirus mRNAs have a common 5' terminal leader sequence encoded at least 10 kb upstream from their main coding regions. Cell. 1977 Sep;12(1):9–21. doi: 10.1016/0092-8674(77)90181-7. [DOI] [PubMed] [Google Scholar]
  25. Lai C. J., Dhar R., Khoury G. Mapping the spliced and unspliced late lytic SV40 RNAs. Cell. 1978 Aug;14(4):971–982. doi: 10.1016/0092-8674(78)90351-3. [DOI] [PubMed] [Google Scholar]
  26. Laskey R. A., Mills A. D. Enhanced autoradiographic detection of 32P and 125I using intensifying screens and hypersensitized film. FEBS Lett. 1977 Oct 15;82(2):314–316. doi: 10.1016/0014-5793(77)80609-1. [DOI] [PubMed] [Google Scholar]
  27. Laub O., Bratosin S., Horowitz M., Aloni Y. The initiation of transcription of SV40 DNA at late time after infection. Virology. 1979 Jan 30;92(2):310–323. doi: 10.1016/0042-6822(79)90136-3. [DOI] [PubMed] [Google Scholar]
  28. Lavi S., Groner Y. 5'-Terminal sequences and coding region of late simian virus 40 mRNAs are derived from noncontiguous segments of the viral genome. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5323–5327. doi: 10.1073/pnas.74.12.5323. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Maitra U., Nakata Y., Hurwitz J. The role of deoxyribonucleic acid in ribonucleic acid synthesis. XIV. A study of the initiation of ribonucleic acid synthesis. J Biol Chem. 1967 Nov 10;242(21):4908–4918. [PubMed] [Google Scholar]
  30. Reddy V. B., Ghosh P. K., Lebowitz P., Weissman S. M. Gaps and duplicated sequences in the leaders of SV40 16S RNA. Nucleic Acids Res. 1978 Nov;5(11):4195–4213. doi: 10.1093/nar/5.11.4195. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Revel M., Groner Y. Post-transcriptional and translational controls of gene expression in eukaryotes. Annu Rev Biochem. 1978;47:1079–1126. doi: 10.1146/annurev.bi.47.070178.005243. [DOI] [PubMed] [Google Scholar]
  32. Shatkin A. J. Capping of eucaryotic mRNAs. Cell. 1976 Dec;9(4 Pt 2):645–653. doi: 10.1016/0092-8674(76)90128-8. [DOI] [PubMed] [Google Scholar]
  33. Silberklang M., Gillum A. M., RajBhandary U. L. The use of nuclease P1 in sequence analysis of end group labeled RNA. Nucleic Acids Res. 1977 Dec;4(12):4091–4108. doi: 10.1093/nar/4.12.4091. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Spencer E., Loring D., Hurwitz J., Monroy G. Enzymatic conversion of 5'-phosphate-terminated RNA to 5'-di- and triphosphate-terminated RNA. Proc Natl Acad Sci U S A. 1978 Oct;75(10):4793–4797. doi: 10.1073/pnas.75.10.4793. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Villarreal L. P., White R. T., Berg P. Mutational alterations within the simian virus 40 leader segment generate altered 16S and 19S mRNA's. J Virol. 1979 Jan;29(1):209–219. doi: 10.1128/jvi.29.1.209-219.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Wei C., Moss B. 5'-Terminal capping of RNA by guanylyltransferase from HeLa cell nuclei. Proc Natl Acad Sci U S A. 1977 Sep;74(9):3758–3761. doi: 10.1073/pnas.74.9.3758. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Weinberg R. A., Warnaar S. O., Winocour E. Isolation and characterization of simian virus 40 ribonucleic acid. J Virol. 1972 Aug;10(2):193–201. doi: 10.1128/jvi.10.2.193-201.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Westphal H., Lai S. P. Quantitative electron microscopy of early adenovirus RNA. J Mol Biol. 1977 Nov 5;116(3):525–548. doi: 10.1016/0022-2836(77)90082-1. [DOI] [PubMed] [Google Scholar]
  39. Winicov I., Perry R. P. Synthesis methylation, and capping of nuclear RNA by a subcellular system. Biochemistry. 1976 Nov 16;15(23):5039–5046. doi: 10.1021/bi00668a014. [DOI] [PubMed] [Google Scholar]
  40. Ziff E. B., Evans R. M. Coincidence of the promoter and capped 5' terminus of RNA from the adenovirus 2 major late transcription unit. Cell. 1978 Dec;15(4):1463–1475. doi: 10.1016/0092-8674(78)90070-3. [DOI] [PubMed] [Google Scholar]

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