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Coronaviruses pp 137-149 | Cite as

Studies into the Mechanism of MHV Transcription

  • Ralph S. Baric
  • Chien Kou Shieh
  • Stephen A. Stohlman
  • Michael M. C. Lai
Chapter
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 218)

Abstract

Coronaviruses are enveloped, plus-polarity RNA viruses which replicate by a unique mode of RNA transcription.1, 2, 3 Previous studies in our laboratory have indicated that RNA recombination occurs at very high frequency during mixed infection with two heterologous strains of MHV4, 5. These data, coupled with the presence of discrete larger leader-containing RNAs which range from 47 to 1000 nucleotides in length in MHV-infected cells,6 suggest that discrete RNA intermediates are synthesized during transcription which may dissociate and reassort between viral RNA templates to generate recombinant viruses by a copy-choice mechanism4. Therefore, the larger leader-containing RNAs in MHV-infected cells may represent functional intermediates of RNA transcription and recombination. In this paper, we have analyzed the origin, structure, and probable mechanism of synthesis of these RNAs. These data provide evidence that MHV RNA transcription pauses at sites corresponding to hairpin loops in the RNA template or product strands and that these RNA intermediates may dissociate and reassociate with the RNA template intermittently during the course of transcription.

Keywords

Small RNAs Hairpin Loop Mouse Hepatitis Virus Hairpin Loop Structure 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. 1.
    M.M.C. Lai and S.A. Stohlman, RNA of mouse hepatitis virus, Journal of Virology 26: 236–242 (1978).PubMedGoogle Scholar
  2. 2.
    L.S. Sturman, Chatacterization of a coronavirus 1. Structural proteins: Effects of preperative conditions on the migration of protein in polyacrylamide gels, Virology 77: 637–649 (1977).PubMedCrossRefGoogle Scholar
  3. 3.
    L.S. Sturman, K.V. Holmes, and J. Behnke, Isolation of coronavirus envelope glycoproteins and interaction with the viral nucleocapsid, Journal of Virology 33: 449–462 (1980).PubMedGoogle Scholar
  4. 4.
    S. Makino, J.G. Keck, S.A. Stohlman, and M.M.C. Lai, High Frequency RNA recombination of murine coronaviruses, Journal of Virology 57: 729–737 (1986).PubMedGoogle Scholar
  5. 5.
    M.M.C. Lai, R.S. Baric, S. Makino, J.G. Keck, J. Egbert, J.L. Leibowitz, and S.A. Stohlman, Recombination between nonsegmented RNA genomes of murine coronaviruses Journal of Virology 56: 449–456 (1985).PubMedGoogle Scholar
  6. 6.
    R.S. Baric, S.A. Stohlman, M.K. Razavi, and M.M.C. Lai, Characterization of leader-related small RNAs in coronavirus-infected cells: Further evidence for leader-primed mechanism of transcription, Virus Res 3: 19–33 (1985).PubMedCrossRefGoogle Scholar
  7. 7.
    R.S. Baric, S.A. Stohlman, and M.M.C. Lai, Characterization of replicative intermediate RNA of mouse hepatitis virus: Presence of leader RNA sequences on nascent chains, Journal of Virology 48: 633–640 (1983).PubMedGoogle Scholar
  8. 8.
    C.K. Shieh, L. Soe, S. Makino, M.F. Chang, S.A. Stohlman, and M.M.C. Lai, The 5′-end sequence of murine coronavirus genome: Implications for multiple fusion sites in leader-primed transcription, (Virology, accepted).Google Scholar
  9. 9.
    J. Armstrong, H. Neiman, S. Smeekens, P. Rottier, and G. Warren, Sequence and topology of a model intracellular membrane protein, E1 glycoprotein, from a coronavirus, Nature 308: 751–752 (1984)PubMedCrossRefGoogle Scholar
  10. 10.
    M.A. Skinner and S.G. Siddell, Coding sequence of coronavirus MHV-JHM mRNA4, Journal of General Virology 66: 593–596 (1985).PubMedCrossRefGoogle Scholar
  11. 11.
    T. Maniatis, E.F. Fritsch, and J. Sambrook, Enzymes used in Molecular Cloning, in “Molecular Cloning — a Laboratory Manual”, pgs 97–148 ed., Cold Spring Harbor Laboratory, New York City (1982).Google Scholar
  12. 12.
    M. Zucker, and P. Stiegler, Optimal computer folding of large RNA sequence using thermodynamics and auxiliary information, Nucleic Acids Res 9: 133–148 (1981).CrossRefGoogle Scholar
  13. 13.
    I. Tinoco, P.N. Borer, B. Dengler, M.D. Levine, O.C. Uhlenbeck, D.M. Crothers, and J. Gralla, Improved estimation of secondary structure in ribonucleic acids, Nature New Biology 246: 40–41 (1973).PubMedGoogle Scholar
  14. 14.
    W. Salser, Globin mRNA sequences: Analysis of base pairing and evolutionary implications, Cold Spring Harbor Svmp. Quant. Biology. 42: 985–1002 (1977).CrossRefGoogle Scholar
  15. 15.
    F.L. Graham, and A.J. Vander Eb, A new technique for the assay of infectivity of human Adenovirus 5 DNA, Virology 52: 456–461.Google Scholar
  16. 16.
    P.S. Thomas, Hybridization of denatured RNA and small DNA fragments transfered to nitrocellulose, PNAS USA 77: 5201–5205 (1980).PubMedCrossRefGoogle Scholar
  17. 17.
    P.R. Mills, C. Dabkin, and F.R. Kramer, Template-determined variable rate of RNA chain elongation, Cell 15: 541–550 (1978).PubMedCrossRefGoogle Scholar
  18. 18.
    C.C. Huang, and J. E. Hearst, Fine mapping of secondary structure of fd phage DNA in the region of the replication origin, Nucleic Acid Res 9: 5587–5599 (1981).PubMedCrossRefGoogle Scholar
  19. 19.
    M. Rosenberg, D. Court, H. Shimatake, C. Brady, and D.L. Wulff, The relationship between function and DNA sequence in an intercistronic region in phage lambda, Nature 272: 414–423 (1978).PubMedCrossRefGoogle Scholar
  20. 20.
    A. Efstratiadis, T. Maniatis, F.C. Kafatos, A. Jeffrey, and J.N. Vournakis, Full length and discrete partial reverse transcripts of globin and chorion mRNAs, Cell 4: 367–378 (1975).PubMedCrossRefGoogle Scholar
  21. 21.
    P.E. Auron, L.D. Weber, and A. Rich, Comparison of transfer ribonucleic acid structures using cobra venom and Sg endonucleases, Biochemistry 21: 4700–4706 (1982).PubMedCrossRefGoogle Scholar
  22. 22.
    F.R. Kramer, and D.R. Mills, Secondary structure formation during RNA synthesis, Nucleic Acids Res 9: 5109–5124 (1981).PubMedCrossRefGoogle Scholar
  23. 23.
    J. Keck, S.A. Stohlman, L. Soe, S. Makino, and M.M.C. Lai, Multiple recombination sites at the 5′-end of murine coronavirus RNA, (Virology accepted).Google Scholar
  24. 24.
    Lyudmila I. Romanova, Vladimir M. Blinov, Elena A. Tolskaya, Ekaterina G. Viktorova, Marina S. Kolesnikova, Elena A. Suseva, and Vadim I. Agol. Virology 155: 202–213 (1986).PubMedCrossRefGoogle Scholar
  25. 25.
    C.C. Huang, N. Hay, and J.M. Bishop, The role of RNA molecules in transduction of the proto-oncogene c-fps, Cell 44: 935–940 (1986).PubMedCrossRefGoogle Scholar
  26. 26.
    E.G. Minkley, and D. Pribnow, Transcription of the early region of bacteriophage T7: Selective initiation with dinucleotides, Journal of Molecular Biology 77: 255–277 (1973).PubMedCrossRefGoogle Scholar
  27. 27.
    O.C. Richards, S.C. Martin, H.G. Jense, and E. Ehrenfeld, The Structure of poliovirus replicative intermediate RNA: Electron Microscope analysis of RNA cross-linked in vivo with psoralen derivitive. Journal of Molecular Biology 173: 324–340 (1984).CrossRefGoogle Scholar
  28. 28.
    S. Makimo, S.A. Stohlman, and M.M.C. Lai, Leader Sequences of murine coronaviruses mRNAs can be freely reassorted: Evidence for the role of free leader RNAs in transcription, PNAS USA 83: 4204–4208 (1986).CrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1987

Authors and Affiliations

  • Ralph S. Baric
    • 1
  • Chien Kou Shieh
    • 2
  • Stephen A. Stohlman
    • 2
  • Michael M. C. Lai
    • 1
  1. 1.School of Public Health, Department of Parasitology and Laboratory PracticeUniversity of North CarolinaChapel HillUSA
  2. 2.School of Medicine, Department of Microbiology and NeurologyUniversity of Southern CaliforniaLos AngelesUSA

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