Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1994 Mar;68(3):1360–1370. doi: 10.1128/jvi.68.3.1360-1370.1994

Targeting of a multicomponent transcription apparatus into assembling vaccinia virus particles requires RAP94, an RNA polymerase-associated protein.

Y Zhang 1, B Y Ahn 1, B Moss 1
PMCID: PMC236590  PMID: 8107201

Abstract

When expression of the vaccinia virus gene encoding RAP94 (a protein that is associated with the viral multisubunit RNA polymerase and confers transcriptional specificity for early promoters) was repressed, the infectious virus yield was reduced by more than 99%. Nevertheless, intermediate- and late-stage viral gene expression and formation of ultrastructurally mature, membrane-enveloped virions occurred under the nonpermissive conditions. The RAP94-deficient particles contained the viral genome, structural proteins, early transcription factor, and certain enzymes but, unlike normal virions, had low or undetectable amounts of the viral RNA polymerase, capping enzyme/termination factor, poly(A) polymerase, DNA-dependent ATPase, RNA helicase, and topoisomerase. The presence of these viral enzymes in the cytoplasm indicated that RAP94 is required for targeting a complex of functionally related proteins involved in the biosynthesis of mRNA.

Full text

PDF
1360

Images in this article

1361Image
on p.1361
1363Image
on p.1363
1364Image
on p.1364
1365Image
on p.1365
1365Image
on p.1365
1366Image
on p.1366
1367Image
on p.1367

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. Glutaredoxin homolog encoded by vaccinia virus is a virion-associated enzyme with thioltransferase and dehydroascorbate reductase activities. Proc Natl Acad Sci U S A. 1992 Aug 1;89(15):7060–7064. doi: 10.1073/pnas.89.15.7060. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Ahn B. Y., Moss B. RNA polymerase-associated transcription specificity factor encoded by vaccinia virus. Proc Natl Acad Sci U S A. 1992 Apr 15;89(8):3536–3540. doi: 10.1073/pnas.89.8.3536. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Banham A. H., Smith G. L. Vaccinia virus gene B1R encodes a 34-kDa serine/threonine protein kinase that localizes in cytoplasmic factories and is packaged into virions. Virology. 1992 Dec;191(2):803–812. doi: 10.1016/0042-6822(92)90256-o. [DOI] [PubMed] [Google Scholar]
  4. Bauer W. R., Ressner E. C., Kates J., Patzke J. V. A DNA nicking-closing enzyme encapsidated in vaccinia virus: partial purification and properties. Proc Natl Acad Sci U S A. 1977 May;74(5):1841–1845. doi: 10.1073/pnas.74.5.1841. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Broyles S. S., Moss B. Identification of the vaccinia virus gene encoding nucleoside triphosphate phosphohydrolase I, a DNA-dependent ATPase. J Virol. 1987 May;61(5):1738–1742. doi: 10.1128/jvi.61.5.1738-1742.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Broyles S. S., Moss B. Sedimentation of an RNA polymerase complex from vaccinia virus that specifically initiates and terminates transcription. Mol Cell Biol. 1987 Jan;7(1):7–14. doi: 10.1128/mcb.7.1.7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Broyles S. S., Yuen L., Shuman S., Moss B. Purification of a factor required for transcription of vaccinia virus early genes. J Biol Chem. 1988 Aug 5;263(22):10754–10760. [PubMed] [Google Scholar]
  8. Condit R. C., Motyczka A., Spizz G. Isolation, characterization, and physical mapping of temperature-sensitive mutants of vaccinia virus. Virology. 1983 Jul 30;128(2):429–443. doi: 10.1016/0042-6822(83)90268-4. [DOI] [PubMed] [Google Scholar]
  9. Dales S., Mosbach E. H. Vaccinia as a model for membrane biogenesis. Virology. 1968 Aug;35(4):564–583. doi: 10.1016/0042-6822(68)90286-9. [DOI] [PubMed] [Google Scholar]
  10. Davison A. J., Moss B. Structure of vaccinia virus late promoters. J Mol Biol. 1989 Dec 20;210(4):771–784. doi: 10.1016/0022-2836(89)90108-3. [DOI] [PubMed] [Google Scholar]
  11. Essani K., Dales S. Biogenesis of vaccinia: evidence for more than 100 polypeptides in the virion. Virology. 1979 Jun;95(2):385–394. doi: 10.1016/0042-6822(79)90493-8. [DOI] [PubMed] [Google Scholar]
  12. Falkner F. G., Moss B. Escherichia coli gpt gene provides dominant selection for vaccinia virus open reading frame expression vectors. J Virol. 1988 Jun;62(6):1849–1854. doi: 10.1128/jvi.62.6.1849-1854.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Falkner F. G., Moss B. Transient dominant selection of recombinant vaccinia viruses. J Virol. 1990 Jun;64(6):3108–3111. doi: 10.1128/jvi.64.6.3108-3111.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Fling S. P., Gregerson D. S. Peptide and protein molecular weight determination by electrophoresis using a high-molarity tris buffer system without urea. Anal Biochem. 1986 May 15;155(1):83–88. doi: 10.1016/0003-2697(86)90228-9. [DOI] [PubMed] [Google Scholar]
  15. Fuerst T. R., Fernandez M. P., Moss B. Transfer of the inducible lac repressor/operator system from Escherichia coli to a vaccinia virus expression vector. Proc Natl Acad Sci U S A. 1989 Apr;86(8):2549–2553. doi: 10.1073/pnas.86.8.2549. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Garcés J., Masternak K., Kunz B., Wittek R. Reactivation of transcription from a vaccinia virus early promoter late in infection. J Virol. 1993 Sep;67(9):5394–5401. doi: 10.1128/jvi.67.9.5394-5401.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Gershon P. D., Ahn B. Y., Garfield M., Moss B. Poly(A) polymerase and a dissociable polyadenylation stimulatory factor encoded by vaccinia virus. Cell. 1991 Sep 20;66(6):1269–1278. doi: 10.1016/0092-8674(91)90048-4. [DOI] [PubMed] [Google Scholar]
  18. Gershon P. D., Moss B. Early transcription factor subunits are encoded by vaccinia virus late genes. Proc Natl Acad Sci U S A. 1990 Jun;87(11):4401–4405. doi: 10.1073/pnas.87.11.4401. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Grimley P. M., Rosenblum E. N., Mims S. J., Moss B. Interruption by Rifampin of an early stage in vaccinia virus morphogenesis: accumulation of membranes which are precursors of virus envelopes. J Virol. 1970 Oct;6(4):519–533. doi: 10.1128/jvi.6.4.519-533.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hagler J., Shuman S. A freeze-frame view of eukaryotic transcription during elongation and capping of nascent mRNA. Science. 1992 Feb 21;255(5047):983–986. doi: 10.1126/science.1546295. [DOI] [PubMed] [Google Scholar]
  21. Horton R. M., Hunt H. D., Ho S. N., Pullen J. K., Pease L. R. Engineering hybrid genes without the use of restriction enzymes: gene splicing by overlap extension. Gene. 1989 Apr 15;77(1):61–68. doi: 10.1016/0378-1119(89)90359-4. [DOI] [PubMed] [Google Scholar]
  22. Jones E. V., Puckett C., Moss B. DNA-dependent RNA polymerase subunits encoded within the vaccinia virus genome. J Virol. 1987 Jun;61(6):1765–1771. doi: 10.1128/jvi.61.6.1765-1771.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Kane E. M., Shuman S. Temperature-sensitive mutations in the vaccinia virus H4 gene encoding a component of the virion RNA polymerase. J Virol. 1992 Oct;66(10):5752–5762. doi: 10.1128/jvi.66.10.5752-5762.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Kates J., Beeson J. Ribonucleic acid synthesis in vaccinia virus. I. The mechanism of synthesis and release of RNA in vaccinia cores. J Mol Biol. 1970 May 28;50(1):1–18. doi: 10.1016/0022-2836(70)90100-2. [DOI] [PubMed] [Google Scholar]
  25. Lin S., Chen W., Broyles S. S. The vaccinia virus B1R gene product is a serine/threonine protein kinase. J Virol. 1992 May;66(5):2717–2723. doi: 10.1128/jvi.66.5.2717-2723.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Martin S. A., Paoletti E., Moss B. Purification of mRNA guanylyltransferase and mRNA (guanine-7-) methyltransferase from vaccinia virions. J Biol Chem. 1975 Dec 25;250(24):9322–9329. [PubMed] [Google Scholar]
  27. Merchlinsky M., Garon C. F., Moss B. Molecular cloning and sequence of the concatemer junction from vaccinia virus replicative DNA. Viral nuclease cleavage sites in cruciform structures. J Mol Biol. 1988 Feb 5;199(3):399–413. doi: 10.1016/0022-2836(88)90613-4. [DOI] [PubMed] [Google Scholar]
  28. Morgan C. The insertion of DNA into vaccinia virus. Science. 1976 Aug 13;193(4253):591–592. doi: 10.1126/science.959819. [DOI] [PubMed] [Google Scholar]
  29. Morgan J. R., Cohen L. K., Roberts B. E. Identification of the DNA sequences encoding the large subunit of the mRNA-capping enzyme of vaccinia virus. J Virol. 1984 Oct;52(1):206–214. doi: 10.1128/jvi.52.1.206-214.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Moss B., Ahn B. Y., Amegadzie B., Gershon P. D., Keck J. G. Cytoplasmic transcription system encoded by vaccinia virus. J Biol Chem. 1991 Jan 25;266(3):1355–1358. [PubMed] [Google Scholar]
  31. Moss B. Regulation of vaccinia virus transcription. Annu Rev Biochem. 1990;59:661–688. doi: 10.1146/annurev.bi.59.070190.003305. [DOI] [PubMed] [Google Scholar]
  32. 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]
  33. Nagaya A., Pogo B. G., Dales S. Biogenesis of vaccinia: separation of early stages from maturation by means of rifampicin. Virology. 1970 Apr;40(4):1039–1051. doi: 10.1016/0042-6822(70)90150-9. [DOI] [PubMed] [Google Scholar]
  34. Niles E. G., Lee-Chen G. J., Shuman S., Moss B., Broyles S. S. Vaccinia virus gene D12L encodes the small subunit of the viral mRNA capping enzyme. Virology. 1989 Oct;172(2):513–522. doi: 10.1016/0042-6822(89)90194-3. [DOI] [PubMed] [Google Scholar]
  35. Paolette E., Rosemond-Hornbeak H., Moss B. Two nucleid acid-dependent nucleoside triphosphate phosphohydrolases from vaccinia virus. Purification and characterization. J Biol Chem. 1974 May 25;249(10):3273–3280. [PubMed] [Google Scholar]
  36. Paoletti E., Moss B. Two nucleic acid-dependent nucleoside triphosphate phosphohydrolases from vaccinia virus. Nucleotide substrate and polynucleotide cofactor specificities. J Biol Chem. 1974 May 25;249(10):3281–3286. [PubMed] [Google Scholar]
  37. Rempel R. E., Traktman P. Vaccinia virus B1 kinase: phenotypic analysis of temperature-sensitive mutants and enzymatic characterization of recombinant proteins. J Virol. 1992 Jul;66(7):4413–4426. doi: 10.1128/jvi.66.7.4413-4426.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Rodriguez J. F., Kahn J. S., Esteban M. Molecular cloning, encoding sequence, and expression of vaccinia virus nucleic acid-dependent nucleoside triphosphatase gene. Proc Natl Acad Sci U S A. 1986 Dec;83(24):9566–9570. doi: 10.1073/pnas.83.24.9566. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Rodriguez J. F., Smith G. L. IPTG-dependent vaccinia virus: identification of a virus protein enabling virion envelopment by Golgi membrane and egress. Nucleic Acids Res. 1990 Sep 25;18(18):5347–5351. doi: 10.1093/nar/18.18.5347. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Roening G., Holowczak J. A. Evidence for the presence of RNA in the purified virions of vaccinia virus. J Virol. 1974 Sep;14(3):704–708. doi: 10.1128/jvi.14.3.704-708.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. 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]
  42. 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]
  43. Rosemond-Hornbeak H., Moss B. Single-stranded deoxyribonucleic acid-specific nuclease from vaccinia virus. Endonucleolytic and exonucleolytic activities. J Biol Chem. 1974 May 25;249(10):3292–3296. [PubMed] [Google Scholar]
  44. Schnierle B. S., Gershon P. D., Moss B. Cap-specific mRNA (nucleoside-O2'-)-methyltransferase and poly(A) polymerase stimulatory activities of vaccinia virus are mediated by a single protein. Proc Natl Acad Sci U S A. 1992 Apr 1;89(7):2897–2901. doi: 10.1073/pnas.89.7.2897. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Shuman S., Golder M., Moss B. Characterization of vaccinia virus DNA topoisomerase I expressed in Escherichia coli. J Biol Chem. 1988 Nov 5;263(31):16401–16407. [PubMed] [Google Scholar]
  46. Shuman S., Hurwitz J. Mechanism of mRNA capping by vaccinia virus guanylyltransferase: characterization of an enzyme--guanylate intermediate. Proc Natl Acad Sci U S A. 1981 Jan;78(1):187–191. doi: 10.1073/pnas.78.1.187. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Shuman S., Moss B. Identification of a vaccinia virus gene encoding a type I DNA topoisomerase. Proc Natl Acad Sci U S A. 1987 Nov;84(21):7478–7482. doi: 10.1073/pnas.84.21.7478. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Shuman S., Moss B. Purification and use of vaccinia virus messenger RNA capping enzyme. Methods Enzymol. 1990;181:170–180. doi: 10.1016/0076-6879(90)81119-f. [DOI] [PubMed] [Google Scholar]
  49. Shuman S. Vaccinia virus RNA helicase: an essential enzyme related to the DE-H family of RNA-dependent NTPases. Proc Natl Acad Sci U S A. 1992 Nov 15;89(22):10935–10939. doi: 10.1073/pnas.89.22.10935. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Sodeik B., Doms R. W., Ericsson M., Hiller G., Machamer C. E., van 't Hof W., van Meer G., Moss B., Griffiths G. Assembly of vaccinia virus: role of the intermediate compartment between the endoplasmic reticulum and the Golgi stacks. J Cell Biol. 1993 May;121(3):521–541. doi: 10.1083/jcb.121.3.521. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Venkatesan S., Moss B. Eukaryotic mRNA capping enzyme-guanylate covalent intermediate. Proc Natl Acad Sci U S A. 1982 Jan;79(2):340–344. doi: 10.1073/pnas.79.2.340. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Wright C. F., Moss B. Identification of factors specific for transcription of the late class of vaccinia virus genes. J Virol. 1989 Oct;63(10):4224–4233. doi: 10.1128/jvi.63.10.4224-4233.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Yuen L., Davison A. J., Moss B. Early promoter-binding factor from vaccinia virions. Proc Natl Acad Sci U S A. 1987 Sep;84(17):6069–6073. doi: 10.1073/pnas.84.17.6069. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Zhang Y. F., Moss B. Inducer-dependent conditional-lethal mutant animal viruses. Proc Natl Acad Sci U S A. 1991 Feb 15;88(4):1511–1515. doi: 10.1073/pnas.88.4.1511. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Zhang Y. F., Moss B. Vaccinia virus morphogenesis is interrupted when expression of the gene encoding an 11-kilodalton phosphorylated protein is prevented by the Escherichia coli lac repressor. J Virol. 1991 Nov;65(11):6101–6110. doi: 10.1128/jvi.65.11.6101-6110.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  56. Zhang Y., Keck J. G., Moss B. Transcription of viral late genes is dependent on expression of the viral intermediate gene G8R in cells infected with an inducible conditional-lethal mutant vaccinia virus. J Virol. 1992 Nov;66(11):6470–6479. doi: 10.1128/jvi.66.11.6470-6479.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  57. Zhang Y., Moss B. Immature viral envelope formation is interrupted at the same stage by lac operator-mediated repression of the vaccinia virus D13L gene and by the drug rifampicin. Virology. 1992 Apr;187(2):643–653. doi: 10.1016/0042-6822(92)90467-4. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES