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. 1990 Feb;87(4):1536–1540. doi: 10.1073/pnas.87.4.1536

Vaccinia virus directs the synthesis of early mRNAs containing 5' poly(A) sequences.

B S Ink 1, D J Pickup 1
PMCID: PMC53510  PMID: 1968253

Abstract

mRNAs transcribed from late promoters of several poxvirus genes contain 5' poly(A) sequences that are not complementary to the viral DNA. In contrast, early mRNAs containing 5' poly(A) sequences have not previously been identified. Modifications to the sequence of the promoter of an early gene of cowpox virus enable this promoter to direct the synthesis of RNAs containing 5' poly(A) sequences. When the sequence 3'-ATTTA-5', which is present at the RNA start-sites of several late promoters, is positioned such that the RNA start-site of the early promoter is at the first thymidylate in this sequence, this early promoter directs the synthesis of early RNAs containing 4-11 adenylates at their 5' ends. When two of the thymidylates in the sequence 3'-ATTTA-5' are removed, the promoter directs the synthesis of early RNAs lacking 5' poly(A) sequences. These results are consistent with the proposal that 5' polyadenylylation of poxvirus RNAs occurs by repetitive transcription of thymidylates in the sequence 3'-ATTTA-5' often present at the sites of transcriptional initiation. In addition, these results demonstrate that 5' polyadenylylation of viral RNAs is not exclusively a late function. The promoter regions of a few early genes of vaccinia virus contain the sequence 3'-ATTTA-5'. Analyses of the transcripts of one of these genes, the D5 gene, indicated that these mRNAs contain 5' poly(A) sequences, suggesting that early mRNAs of a subset of viral genes contain 5' poly(A) sequences.

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Selected References

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

  1. Ahn B. Y., Moss B. Capped poly(A) leaders of variable lengths at the 5' ends of vaccinia virus late mRNAs. J Virol. 1989 Jan;63(1):226–232. doi: 10.1128/jvi.63.1.226-232.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Berk A. J., Sharp P. A. Sizing and mapping of early adenovirus mRNAs by gel electrophoresis of S1 endonuclease-digested hybrids. Cell. 1977 Nov;12(3):721–732. doi: 10.1016/0092-8674(77)90272-0. [DOI] [PubMed] [Google Scholar]
  3. Bertholet C., Drillien R., Wittek R. One hundred base pairs of 5' flanking sequence of a vaccinia virus late gene are sufficient to temporally regulate late transcription. Proc Natl Acad Sci U S A. 1985 Apr;82(7):2096–2100. doi: 10.1073/pnas.82.7.2096. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bertholet C., Stocco P., Van Meir E., Wittek R. Functional analysis of the 5' flanking sequence of a vaccinia virus late gene. EMBO J. 1986 Aug;5(8):1951–1957. doi: 10.1002/j.1460-2075.1986.tb04449.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bertholet C., Van Meir E., ten Heggeler-Bordier B., Wittek R. Vaccinia virus produces late mRNAs by discontinuous synthesis. Cell. 1987 Jul 17;50(2):153–162. doi: 10.1016/0092-8674(87)90211-X. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Broyles S. S., Moss B. Homology between RNA polymerases of poxviruses, prokaryotes, and eukaryotes: nucleotide sequence and transcriptional analysis of vaccinia virus genes encoding 147-kDa and 22-kDa subunits. Proc Natl Acad Sci U S A. 1986 May;83(10):3141–3145. doi: 10.1073/pnas.83.10.3141. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Carter P., Bedouelle H., Winter G. Improved oligonucleotide site-directed mutagenesis using M13 vectors. Nucleic Acids Res. 1985 Jun 25;13(12):4431–4443. doi: 10.1093/nar/13.12.4431. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cochran M. A., Puckett C., Moss B. In vitro mutagenesis of the promoter region for a vaccinia virus gene: evidence for tandem early and late regulatory signals. J Virol. 1985 Apr;54(1):30–37. doi: 10.1128/jvi.54.1.30-37.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Coupar B. E., Boyle D. B., Both G. W. Effect of in vitro mutations in a vaccinia virus early promoter region monitored by herpes simplex virus thymidine kinase expression in recombinant vaccinia virus. J Gen Virol. 1987 Sep;68(Pt 9):2299–2309. doi: 10.1099/0022-1317-68-9-2299. [DOI] [PubMed] [Google Scholar]
  10. Earl P. L., Jones E. V., Moss B. Homology between DNA polymerases of poxviruses, herpesviruses, and adenoviruses: nucleotide sequence of the vaccinia virus DNA polymerase gene. Proc Natl Acad Sci U S A. 1986 Jun;83(11):3659–3663. doi: 10.1073/pnas.83.11.3659. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Eisen H. RNA editing: who's on first? Cell. 1988 May 6;53(3):331–332. doi: 10.1016/0092-8674(88)90150-x. [DOI] [PubMed] [Google Scholar]
  12. Ghosh P. K., Reddy V. B., Piatak M., Lebowitz P., Weissman S. M. Determination of RNA sequences by primer directed synthesis and sequencing of their cDNA transcripts. Methods Enzymol. 1980;65(1):580–595. doi: 10.1016/s0076-6879(80)65061-7. [DOI] [PubMed] [Google Scholar]
  13. Hirt P., Hiller G., Wittek R. Localization and fine structure of a vaccinia virus gene encoding an envelope antigen. J Virol. 1986 Jun;58(3):757–764. doi: 10.1128/jvi.58.3.757-764.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Holowczak J. A., Joklik W. K. Studies on the structural proteins of vaccinia virus. I. Structural proteins of virions and cores. Virology. 1967 Dec;33(4):717–725. doi: 10.1016/0042-6822(67)90072-4. [DOI] [PubMed] [Google Scholar]
  15. Hänggi M., Bannwarth W., Stunnenberg H. G. Conserved TAAAT motif in vaccinia virus late promoters: overlapping TATA box and site of transcription initiation. EMBO J. 1986 May;5(5):1071–1076. doi: 10.1002/j.1460-2075.1986.tb04324.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Ink B. S., Pickup D. J. Transcription of a poxvirus early gene is regulated both by a short promoter element and by a transcriptional termination signal controlling transcriptional interference. J Virol. 1989 Nov;63(11):4632–4644. doi: 10.1128/jvi.63.11.4632-4644.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kassavetis G. A., Zentner P. G., Geiduschek E. P. Transcription at bacteriophage T4 variant late promoters. An application of a newly devised promoter-mapping method involving RNA chain retraction. J Biol Chem. 1986 Oct 25;261(30):14256–14265. [PubMed] [Google Scholar]
  18. Lee-Chen G. J., Bourgeois N., Davidson K., Condit R. C., Niles E. G. Structure of the transcription initiation and termination sequences of seven early genes in the vaccinia virus HindIII D fragment. Virology. 1988 Mar;163(1):64–79. doi: 10.1016/0042-6822(88)90234-6. [DOI] [PubMed] [Google Scholar]
  19. Lee-Chen G. J., Niles E. G. Map positions of the 5' ends of eight mRNAs synthesized from the late genes in the vaccinia virus HindIII D fragment. Virology. 1988 Mar;163(1):80–92. doi: 10.1016/0042-6822(88)90235-8. [DOI] [PubMed] [Google Scholar]
  20. Macaulay C., McFadden G. Tumorigenic poxviruses: characterization of an early promoter from Shope fibroma virus. Virology. 1989 Sep;172(1):237–246. doi: 10.1016/0042-6822(89)90125-6. [DOI] [PubMed] [Google Scholar]
  21. Mackett M., Smith G. L., Moss B. General method for production and selection of infectious vaccinia virus recombinants expressing foreign genes. J Virol. 1984 Mar;49(3):857–864. doi: 10.1128/jvi.49.3.857-864.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Niles E. G., Condit R. C., Caro P., Davidson K., Matusick L., Seto J. Nucleotide sequence and genetic map of the 16-kb vaccinia virus HindIII D fragment. Virology. 1986 Aug;153(1):96–112. doi: 10.1016/0042-6822(86)90011-5. [DOI] [PubMed] [Google Scholar]
  23. Patel D. D., Pickup D. J. Messenger RNAs of a strongly-expressed late gene of cowpox virus contain 5'-terminal poly(A) sequences. EMBO J. 1987 Dec 1;6(12):3787–3794. doi: 10.1002/j.1460-2075.1987.tb02714.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Pickup D. J., Ink B. S., Hu W., Ray C. A., Joklik W. K. Hemorrhage in lesions caused by cowpox virus is induced by a viral protein that is related to plasma protein inhibitors of serine proteases. Proc Natl Acad Sci U S A. 1986 Oct;83(20):7698–7702. doi: 10.1073/pnas.83.20.7698. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Rice A. P., Roberts B. E. Vaccinia virus induces cellular mRNA degradation. J Virol. 1983 Sep;47(3):529–539. doi: 10.1128/jvi.47.3.529-539.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Rosel J. L., Earl P. L., Weir J. P., Moss B. Conserved TAAATG sequence at the transcriptional and translational initiation sites of vaccinia virus late genes deduced by structural and functional analysis of the HindIII H genome fragment. J Virol. 1986 Nov;60(2):436–449. doi: 10.1128/jvi.60.2.436-449.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Roseman N. A., Hruby D. E. Nucleotide sequence and transcript organization of a region of the vaccinia virus genome which encodes a constitutively expressed gene required for DNA replication. J Virol. 1987 May;61(5):1398–1406. doi: 10.1128/jvi.61.5.1398-1406.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Sachs A. B., Davis R. W. The poly(A) binding protein is required for poly(A) shortening and 60S ribosomal subunit-dependent translation initiation. Cell. 1989 Sep 8;58(5):857–867. doi: 10.1016/0092-8674(89)90938-0. [DOI] [PubMed] [Google Scholar]
  29. Schwer B., Stunnenberg H. G. Vaccinia virus late transcripts generated in vitro have a poly(A) head. EMBO J. 1988 Apr;7(4):1183–1190. doi: 10.1002/j.1460-2075.1988.tb02929.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Schwer B., Visca P., Vos J. C., Stunnenberg H. G. Discontinuous transcription or RNA processing of vaccinia virus late messengers results in a 5' poly(A) leader. Cell. 1987 Jul 17;50(2):163–169. doi: 10.1016/0092-8674(87)90212-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Shenk T. E., Rhodes C., Rigby P. W., Berg P. Biochemical method for mapping mutational alterations in DNA with S1 nuclease: the location of deletions and temperature-sensitive mutations in simian virus 40. Proc Natl Acad Sci U S A. 1975 Mar;72(3):989–993. doi: 10.1073/pnas.72.3.989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Venkatesan S., Baroudy B. M., Moss B. Distinctive nucleotide sequences adjacent to multiple initiation and termination sites of an early vaccinia virus gene. Cell. 1981 Sep;25(3):805–813. doi: 10.1016/0092-8674(81)90188-4. [DOI] [PubMed] [Google Scholar]
  33. Venkatesan S., Moss B. In vitro transcription of the inverted terminal repetition of the vaccinia virus genome: correspondence of initiation and cap sites. J Virol. 1981 Feb;37(2):738–747. doi: 10.1128/jvi.37.2.738-747.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Weir J. P., Moss B. Determination of the promoter region of an early vaccinia virus gene encoding thymidine kinase. Virology. 1987 May;158(1):206–210. doi: 10.1016/0042-6822(87)90254-6. [DOI] [PubMed] [Google Scholar]
  35. Weir J. P., Moss B. Determination of the transcriptional regulatory region of a vaccinia virus late gene. J Virol. 1987 Jan;61(1):75–80. doi: 10.1128/jvi.61.1.75-80.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Weir J. P., Moss B. Nucleotide sequence of the vaccinia virus thymidine kinase gene and the nature of spontaneous frameshift mutations. J Virol. 1983 May;46(2):530–537. doi: 10.1128/jvi.46.2.530-537.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Weir J. P., Moss B. Regulation of expression and nucleotide sequence of a late vaccinia virus gene. J Virol. 1984 Sep;51(3):662–669. doi: 10.1128/jvi.51.3.662-669.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Wright C. F., Moss B. In vitro synthesis of vaccinia virus late mRNA containing a 5' poly(A) leader sequence. Proc Natl Acad Sci U S A. 1987 Dec;84(24):8883–8887. doi: 10.1073/pnas.84.24.8883. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. de Magistris L., Stunnenberg H. G. Cis-acting sequences affecting the length of the poly(A) head of vaccinia virus late transcripts. Nucleic Acids Res. 1988 Apr 25;16(8):3141–3156. doi: 10.1093/nar/16.8.3141. [DOI] [PMC free article] [PubMed] [Google Scholar]

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