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. 1975 Jul;16(1):34–42. doi: 10.1128/jvi.16.1.34-42.1975

Inhibition of host protein synthesis by vaccinia virus: fate of cell mRNA and synthesis of small poly (A)-rich polyribonucleotides in the presence of actinomycin D.

H Rosemond-Hornbeak, B Moss
PMCID: PMC354629  PMID: 1133878

Abstract

Purified vaccinia virus rapidly inhibited HeLa cell protein synthesis in the presence of actinomycin D. Under these conditions host polyribosomes were extensively degraded but the mRNA was stable as indicated by a greater than 90% recovery of prelabeled polyadenylylated RNA. Although actinomycin D prevented the synthesis of host mRNA and poly(A) in uninfected cells, incorporation of adenosine into poly(A) was inhibited by less than 50% in infected cells. Further analysis indicated that there was little or no normal size viral mRNA but that a unique class of small poly(A)-rich RNA was made in the presence of actinomycin D. From measurements of the RNase resistance and base composition of the RNA, approximately 40% of the nucleotide sequence was estimated to be poly(A). The poly(A)-rich RNA was found associated with small polyribosomes and monoribosomes that were inactive in protein synthesis. It was suggested that the poly(A) segment of the RNA is formed by the poly(A) polymerase previously found in vaccinia virus cores and that the inactive RNA, by competing with host mRNA, may contribute to the virus-mediated inhibition of host protein synthesis observed in the presence of actinomycin D.

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Selected References

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

  1. Adesnik M., Salditt M., Thomas W., Darnell J. E. Evidence that all messenger RNA molecules (except histone messenger RNA) contain Poly (A) sequences and that the Poly(A) has a nuclear function. J Mol Biol. 1972 Oct 28;71(1):21–30. doi: 10.1016/0022-2836(72)90397-x. [DOI] [PubMed] [Google Scholar]
  2. BECKER Y., JOKLIK W. K. MESSENGER RNA IN CELLS INFECTED WITH VACCINIA VIRUS. Proc Natl Acad Sci U S A. 1964 Apr;51:577–585. doi: 10.1073/pnas.51.4.577. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Colby D. S., Finnerty V., Lucas-Lenard J. Fate of mRNA of L-cells infected with mengovirus. J Virol. 1974 Apr;13(4):858–869. doi: 10.1128/jvi.13.4.858-869.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Esteban M., Metz D. H. Early virus protein synthesis in vaccinia virus-infected cells. J Gen Virol. 1973 May;19(2):201–206. doi: 10.1099/0022-1317-19-2-201. [DOI] [PubMed] [Google Scholar]
  5. Kates J., Beeson J. Ribonucleic acid synthesis in vaccinia virus. II. Synthesis of polyriboadenylic acid. J Mol Biol. 1970 May 28;50(1):19–33. doi: 10.1016/0022-2836(70)90101-4. [DOI] [PubMed] [Google Scholar]
  6. Katz E., Moss B. Vaccinia virus structural polypeptide derived from a high-molecular-weight precursor: formation and integration into virus particles. J Virol. 1970 Dec;6(6):717–726. doi: 10.1128/jvi.6.6.717-726.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Koschel K. Poliovirus infection and poly(A) sequences of cytoplasmic cellular RNA. J Virol. 1974 May;13(5):1061–1066. doi: 10.1128/jvi.13.5.1061-1066.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Lindberg U., Persson T. Isolation of mRNA from KB-cells by affinity chromatography on polyuridylic acid covalently linked to Sepharose. Eur J Biochem. 1972 Dec 4;31(2):246–254. doi: 10.1111/j.1432-1033.1972.tb02527.x. [DOI] [PubMed] [Google Scholar]
  9. Moss B. Inhibition of HeLa cell protein synthesis by the vaccinia virion. J Virol. 1968 Oct;2(10):1028–1037. doi: 10.1128/jvi.2.10.1028-1037.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Moss B., Rosenblum E. N., Paoletti E. Polyadenylate polymerase from vaccinia virions. Nat New Biol. 1973 Sep 12;245(141):59–63. doi: 10.1038/newbio245059a0. [DOI] [PubMed] [Google Scholar]
  11. Moss B., Rosenblum E. N. Vaccinia virus polyriboadenylate polymerase: convalent linkage of the product with polyribonucleotide and polydeoxyribonucleotide primers. J Virol. 1974 Jul;14(1):86–98. doi: 10.1128/jvi.14.1.86-98.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. Perry R. P., La Torre J., Kelley D. E., Greenberg J. R. On the lability of poly(A) sequences during extraction of messenger RNA from polyribosomes. Biochim Biophys Acta. 1972 Mar 14;262(2):220–226. doi: 10.1016/0005-2787(72)90236-5. [DOI] [PubMed] [Google Scholar]
  14. Randerath K. Separation of the constituent nucleotides of nucleic acids on ion-exchange thin-layers. Experientia. 1964 Jul 15;20(7):406–407. doi: 10.1007/BF02147995. [DOI] [PubMed] [Google Scholar]
  15. SALZMAN N. P., SHATKIN A. J., SEBRING E. D. THE SYNTHESIS OF A DNA-LIKE RNA IN THE CYTOPLASM OF HELA CELLS INFECTED WITH VACCINIA VIRUS. J Mol Biol. 1964 Mar;8:405–416. doi: 10.1016/s0022-2836(64)80204-7. [DOI] [PubMed] [Google Scholar]
  16. SHATKIN A. J. ACTINOMYCIN D AND VACCINIA VIRUS INFECTION OF HELA CELLS. Nature. 1963 Jul 27;199:357–358. doi: 10.1038/199357a0. [DOI] [PubMed] [Google Scholar]
  17. Salzman N. P., Sebring E. D. Sequential formation of vaccinia virus proteins and viral deoxyribonucleic acid replication. J Virol. 1967 Feb;1(1):16–23. doi: 10.1128/jvi.1.1.16-23.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Tibbetts C., Johansson K., Philipson L. Hydroxyapatite chromatography and formamide denaturation of adenovirus DNA. J Virol. 1973 Aug;12(2):218–225. doi: 10.1128/jvi.12.2.218-225.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Wagner A. F., Bugianesi R. L., Shen T. Y. Preparation of sepharose-bound poly (rI:rC). Biochem Biophys Res Commun. 1971 Oct 1;45(1):184–189. doi: 10.1016/0006-291x(71)90067-2. [DOI] [PubMed] [Google Scholar]

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