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. 1987 Mar;61(3):639–645. doi: 10.1128/jvi.61.3.639-645.1987

Noncoordinate regulation of a vaccinia virus late gene cluster.

S L Weinrich, D E Hruby
PMCID: PMC254001  PMID: 3806794

Abstract

Identification of a tightly spaced and tandemly oriented late gene cluster within the central conserved region of the vaccinia virus genome suggested the possibility of coordinate regulation of the genes within this domain (S.L. Weinrich and D.E. Hruby, Nucleic Acids Res. 14:3003-3016, 1986). To test this hypothesis, the steady-state levels of transcripts derived from the individual late genes were examined. Cytoplasmic RNA was isolated from infected cells at hourly intervals throughout infection and was used in concert with 5' S1 nuclease mapping procedures to detect transcripts from specific late genes. Among the set of six closely linked late genes, marked differences were observed in both the levels of transcription and the kinetic patterns of expression, providing direct evidence for the existence of differentially regulated gene subsets within the late gene class. Furthermore, these experiments identified one of the genes (encoding a 33,000-molecular-weight polypeptide) as being expressed both early and late postinfection. Interestingly, although transcripts from the constitutively expressed gene were initiated at the same start sites throughout infection, a discrete terminus for these transcripts was detected only at early times. These data suggest that the lack of cis-acting termination signals is not the reason for the late gene transcript heterogeneity observed in vaccinia virus-infected cells.

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Selected References

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

  1. Barbosa E., Moss B. mRNA(nucleoside-2'-)-methyltransferase from vaccinia virus. Characteristics and substrate specificity. J Biol Chem. 1978 Nov 10;253(21):7698–7702. [PubMed] [Google Scholar]
  2. Baroudy B. M., Moss B. Purification and characterization of a DNA-dependent RNA polymerase from vaccinia virions. J Biol Chem. 1980 May 10;255(9):4372–4380. [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. Bossart W., Nuss D. L., Paoletti E. Effect of UV irradiation on the expression of vaccinia virus gene products synthesized in a cell-free system coupling transcription and translation. J Virol. 1978 Jun;26(3):673–680. doi: 10.1128/jvi.26.3.673-680.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chipchase M., Schwendimann F., Wyler R. A map of the late proteins of vaccinia virus. Virology. 1980 Aug;105(1):261–264. doi: 10.1016/0042-6822(80)90176-2. [DOI] [PubMed] [Google Scholar]
  6. Cochran M. A., Mackett M., Moss B. Eukaryotic transient expression system dependent on transcription factors and regulatory DNA sequences of vaccinia virus. Proc Natl Acad Sci U S A. 1985 Jan;82(1):19–23. doi: 10.1073/pnas.82.1.19. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. 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]
  8. Cooper J. A., Moss B. Translation of specific vaccinia virus RNAs purified as RNA-DNA hybrids on potassium iodide gradients. Nucleic Acids Res. 1979 Aug 10;6(11):3599–3612. doi: 10.1093/nar/6.11.3599. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Cooper J. A., Wittek R., Moss B. Extension of the transcriptional and translational map of the left end of the vaccinia virus genome to 21 kilobase pairs. J Virol. 1981 Sep;39(3):733–745. doi: 10.1128/jvi.39.3.733-745.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Cooper J. A., Wittek R., Moss B. Hybridization selection and cell-free translation of mRNA's encoded within the inverted terminal repetition of the vaccinia virus genome. J Virol. 1981 Jan;37(1):284–294. doi: 10.1128/jvi.37.1.284-294.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Golini F., Kates J. R. Transcriptional and translational analysis of a strongly expressed early region of the vaccinia virus genome. J Virol. 1984 Feb;49(2):459–470. doi: 10.1128/jvi.49.2.459-470.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hruby D. E., Guarino L. A., Kates J. R. Vaccinia virus replication. I. Requirement for the host-cell nucleus. J Virol. 1979 Feb;29(2):705–715. doi: 10.1128/jvi.29.2.705-715.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hruby D. E., Lynn D. L., Condit R. C., Kates J. R. Cellular differences in the molecular mechanisms of vaccinia virus host range restriction. J Gen Virol. 1980 Apr;47(2):485–488. doi: 10.1099/0022-1317-47-2-485. [DOI] [PubMed] [Google Scholar]
  14. Isle H. B., Venkatesan S., Moss B. Cell-free translation of early and late mRNAs selected by hybridization to cloned DNA fragments derived from the left 14 million to 72 million daltons of the vaccinia virus genome. Virology. 1981 Jul 15;112(1):306–317. doi: 10.1016/0042-6822(81)90636-x. [DOI] [PubMed] [Google Scholar]
  15. Kaverin N. V., Varich N. L., Surgay V. V., Chernos V. I. A quantitative estimation of poxvirus genome fraction transcribed as "early" and "late" mRNA. Virology. 1975 May;65(1):112–119. doi: 10.1016/0042-6822(75)90011-2. [DOI] [PubMed] [Google Scholar]
  16. Mahr A., Roberts B. E. Arrangement of late RNAs transcribed from a 7.1-kilobase EcoRI vaccinia virus DNA fragment. J Virol. 1984 Feb;49(2):510–520. doi: 10.1128/jvi.49.2.510-520.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Mahr A., Roberts B. E. Organization of six early transcripts synthesized from a vaccinia virus EcoRI DNA fragment. J Virol. 1984 Feb;49(2):497–509. doi: 10.1128/jvi.49.2.497-509.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. 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]
  19. Morgan J. R., Roberts B. E. Organization of RNA transcripts from a vaccinia virus early gene cluster. J Virol. 1984 Aug;51(2):283–297. doi: 10.1128/jvi.51.2.283-297.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Moss B., Rosenblum E. N., Gershowitz A. Characterization of a polyriboadenylate polymerase from vaccinia virions. J Biol Chem. 1975 Jun 25;250(12):4722–4729. [PubMed] [Google Scholar]
  21. Moss B., Salzman N. P. Sequential protein synthesis following vaccinia virus infection. J Virol. 1968 Oct;2(10):1016–1027. doi: 10.1128/jvi.2.10.1016-1027.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Nevins J. R., Joklik W. K. Isolation and properties of the vaccinia virus DNA-dependent RNA polymerase. J Biol Chem. 1977 Oct 10;252(19):6930–6938. [PubMed] [Google Scholar]
  23. Oda K. I., Joklik W. K. Hybridization and sedimentation studies on "early" and "late" vaccinia messenger RNA. J Mol Biol. 1967 Aug 14;27(3):395–419. doi: 10.1016/0022-2836(67)90047-2. [DOI] [PubMed] [Google Scholar]
  24. Paoletti E., Grady L. J. Transcriptional complexity of vaccinia virus in vivo and in vitro. J Virol. 1977 Sep;23(3):608–615. doi: 10.1128/jvi.23.3.608-615.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Pelham H. R., Jackson R. J. An efficient mRNA-dependent translation system from reticulocyte lysates. Eur J Biochem. 1976 Aug 1;67(1):247–256. doi: 10.1111/j.1432-1033.1976.tb10656.x. [DOI] [PubMed] [Google Scholar]
  26. Pelham H. R. Use of coupled transcription and translation to study mRNA production by vaccinia cores. Nature. 1977 Oct 6;269(5628):532–534. doi: 10.1038/269532a0. [DOI] [PubMed] [Google Scholar]
  27. Pennington T. H. Vaccinia virus polypeptide synthesis: sequential appearance and stability of pre- and post-replicative polypeptides. J Gen Virol. 1974 Dec;25(3):433–444. doi: 10.1099/0022-1317-25-3-433. [DOI] [PubMed] [Google Scholar]
  28. Plucienniczak A., Schroeder E., Zettlmeissl G., Streeck R. E. Nucleotide sequence of a cluster of early and late genes in a conserved segment of the vaccinia virus genome. Nucleic Acids Res. 1985 Feb 11;13(3):985–998. doi: 10.1093/nar/13.3.985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. 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]
  30. Rohrmann G., Moss B. Transcription of vaccinia virus early genes by a template-dependent soluble extract of purified virions. J Virol. 1985 Nov;56(2):349–355. doi: 10.1128/jvi.56.2.349-355.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Rosel J., Moss B. Transcriptional and translational mapping and nucleotide sequence analysis of a vaccinia virus gene encoding the precursor of the major core polypeptide 4b. J Virol. 1985 Dec;56(3):830–838. doi: 10.1128/jvi.56.3.830-838.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Sebring E. D., Salzman N. P. Metabolic properties of early and late vaccinia virus messenger ribonucleic acid. J Virol. 1967 Jun;1(3):550–558. doi: 10.1128/jvi.1.3.550-558.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Spencer E., Shuman S., Hurwitz J. Purification and properties of vaccinia virus DNA-dependent RNA polymerase. J Biol Chem. 1980 Jun 10;255(11):5388–5395. [PubMed] [Google Scholar]
  34. 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]
  35. Weinrich S. L., Hruby D. E. A tandemly-oriented late gene cluster within the vaccinia virus genome. Nucleic Acids Res. 1986 Apr 11;14(7):3003–3016. doi: 10.1093/nar/14.7.3003. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Weinrich S. L., Niles E. G., Hruby D. E. Transcriptional and translational analysis of the vaccinia virus late gene L65. J Virol. 1985 Aug;55(2):450–457. doi: 10.1128/jvi.55.2.450-457.1985. [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. Wittek R., Cooper J. A., Moss B. Transcriptional and translational mapping of a 6.6-kilobase-pair DNA fragment containing the junction of the terminal repetition and unique sequence at the left end of the vaccinia virus genome. J Virol. 1981 Sep;39(3):722–732. doi: 10.1128/jvi.39.3.722-732.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Wittek R., Hänggi M., Hiller G. Mapping of a gene coding for a major late structural polypeptide on the vaccinia virus genome. J Virol. 1984 Feb;49(2):371–378. doi: 10.1128/jvi.49.2.371-378.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Wittek R., Richner B., Hiller G. Mapping of the genes coding for the two major vaccinia virus core polypeptides. Nucleic Acids Res. 1984 Jun 25;12(12):4835–4848. doi: 10.1093/nar/12.12.4835. [DOI] [PMC free article] [PubMed] [Google Scholar]

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