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Studies on the Mechanism of RNA Synthesis of a Murine Coronavirus

  • Michael M. C. Lai
  • Ralph S. Baric
  • Peter R. Brayton
  • Stephen A. Stohlman
Chapter
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 173)

Abstract

The mechanism of viral RNA replication in mouse hepatitis virus (MHV)-infected cells was studied by oligonucleotide mapping of every mRNA. We discovered that an oligonucleotide. No. 10, was localized at the 5′-end of every mRNA, and was not colinear with the sequences of the virion genomic RNA. This result indicates that all of the mRNAs contain a leader sequence which is joined to the body sequences of the mRNAs. We have also studied the structure of the replicative intermediate (RI) RNA in the MHV-infected cells. This RI RNA consists of a single species corresponding to the MHV genomic RNA. No subgenomic RI structures were detected. Furthermore, the nascent RNA chains in the RI structure contained the leader sequences, suggesting that the leader RNA was not added to the mRNA post-transciptionally, but rather, it was probably synthesized independently and then used as a primer for the synthesis of mRNAs. We have also shown that the poly (A) sequences in the MHV genome were transcribed from the poly (U) sequences present in the negative-strand template. The RNA polymerases involved in the MHV RNA synthesis were also characterized. The early polymerase synthesizes a single negative-stranded, full-length RNA. The late polymerases could be separated into two activities, one synthesizing positive-stranded genomic RNA, and the other synthesizing genomic as well as subgenomic RNAs. Thus, the replication and transcription functions of MHV could probably be separated. A plausible model of MHV replication is presented.

Keywords

Membrane Complex Replicative Form Replicative Intermediate Mouse Hepatitis Virus Murine Coronavirus 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

References

  1. Brayton, P.R., Lai, M.M.C., Patton, C.D and Stohlman, S.A. 1982. Characterization of two RNA polymerase activities induced by mouse hepatitis virus. J. Virol. 42:847–853.PubMedGoogle Scholar
  2. Colonno, R.J. and A.K. Banerjee. 1976. A unique RNA species involved in initiation of vesicular stomatitis virus RNA transcription. Cell 8:197–204.PubMedCrossRefGoogle Scholar
  3. Jacobs, L., W.J.M. Spaan, M.C. Horzinek and B.A.M. Van der Zeijst. 1981. Synthesis of subgenomic mRNAs of mouse hepatitis virus is initiated independently:evidence from UV transcription mapping. J. Virol. 39:401–406.PubMedGoogle Scholar
  4. Krug, R.M. 1981. Priming of influenza viral RNA transcription by capped heterologous RNAs. Curr. Topics in Microb. Immunol. 93:125–149.CrossRefGoogle Scholar
  5. Lai, M.M.C., Brayton, P.R., Armen, R.C., Patton, C.D., Pugh, C., and S.A. Stohlman. 1981. Mouse hepatitis virus A59: Messenger RNA structure and genetic localization of the sequence divergence from a hepatotropic strain MHV-3. J. Virol. 39:823–834.PubMedGoogle Scholar
  6. Lai, M.M.C., S.S.F. Hu and P.K. Vogt. 1979. Avian erythroblastosis virus: Transformation-specific sequences form a contiguous segment of 3.25 kb located in the middle of the 6-kb genome. Virology 97:366–377.PubMedCrossRefGoogle Scholar
  7. Lai, M.M.C., Patton, C.D., and S.A. Stohlman. 1982a. Further characterization of mRNAs of mouse hepatitis virus: presence of common 5′-end nucleotides. J. Virol. 41:557–565.Google Scholar
  8. Lai, M.M.C., C.D. Patton and S.A. Stohlman. 1982b. Replication of mouse hepatitis virus: negative-stranded RNA and replicative form RNA are of genome length. J. Virol. 44:487–492.Google Scholar
  9. Lai, M.M.C. and S.A. Stohlman. 1978. The RNA of mouse hepatitis virus. J. Virol. 271:236–242.Google Scholar
  10. Lai, M.M.C. and Stohlman, S.A. 1981. Comparative analysis of RNA genome of mouse hepatitis virus. J. Virol. 38:661–670.PubMedGoogle Scholar
  11. Leibowitz, J.L., S.R. Weiss, E. Paavola and C.L. Bond. 1982. Cell-free translation of murine coronavirus RNA. J. Virol. 43:905–913.PubMedGoogle Scholar
  12. Leibowitz, J.L., K.C. Wilhelmsen and C.W. Bond. 1981. The virus-specific intracellular RNA species of two murine coronaviruses: MHV-A59 and MHV-JHM. Virology 114:29–51.CrossRefGoogle Scholar
  13. Mahy, B.W.J., S. Siddell, H. Wege and V. ter Meulen. 1983. RNA-dependent RNA polymerase activity in murine coronavirus-infected cell. J. Gen. Virol. 64:103–111.PubMedCrossRefGoogle Scholar
  14. Spaan, W.J.M., P.J.M. Rottier, M.C. Horzinek and B.A.M. van der Zeijst. 1981. Isolation and identification of virus-specific mRNAs in cells infected with mouse hepatitis virus (MHV-A59). Virology 108:424–434.PubMedCrossRefGoogle Scholar
  15. Spector, D.H. and D. Baltimore. 1975. Polyadenylic acid on poliovirus RNA. II. Poly A on intracellular RNAs. J. Virol. 15:1418–1431.PubMedGoogle Scholar
  16. Sturman, L.S. and K.V. Homes. 1983. The molecular biology of coronaviruses. Adv. in Virus Research (in press).Google Scholar

Copyright information

© Plenum Press, New York 1984

Authors and Affiliations

  • Michael M. C. Lai
    • 1
  • Ralph S. Baric
    • 1
  • Peter R. Brayton
    • 1
  • Stephen A. Stohlman
    • 1
  1. 1.School of MedicineUniversity of Southern CaliforniaLos AngelesUSA

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