Advertisement

Protein-protein interactions and glycerophospholipids in bromovirus and nodavirus RNA replication

  • P. Ahlquist
  • S.-X. Wu
  • P. Kaesberg
  • C. C. Kao
  • R. Quadt
  • W. Dejong
  • R. Hershberger
Conference paper
Part of the Archives of Virology Supplementum book series (ARCHIVES SUPPL, volume 9)

Summary

The plant bromoviruses and animal nodaviruses are distinct groups of positive strand RNA viruses that have proven to be useful models for RNA replication studies. Bromoviruses encode two large proteins required for RNA replication: 1a contains domains implicated in helicase and capping functions, and 2a contains a central polymerase-like domain. Using immunoprecipitation and far-western blotting, we have now shown that la and 2a form a specific complex in vitro and have mapped the interacting domains. Molecular genetic data implicate the 1a–2a complex in RNA replication and suggest that it supports coordinate action of the putative helicase, polymerase, and capping domains. The locations of the interacting la and 2a domains have implications for replication models and the evolution of virus genomes bearing homologous replication genes in fused vs. divided forms. For the nodavirus Flock house virus (FHV), a true RNA replicase has been isolated that carries out complete, highly active replication of added FHV RNA, producing newly synthesized positive strand RNA in pre-dominantly ssRNA form. Positive strand RNA synthesis in this FHV cell-free system is strongly dependent on the addition of any of several glycerophospholipids. Positive strand RNA synthesis depends on the complete glycerophospholipid structure, including the polar head group and diacyl glycerol lipid portion, and is strongly influenced by acyl chain length.

Keywords

Positive Strand Brome Mosaic Virus Cowpea Chlorotic Mottle Virus Flock House Virus dsRNA Product 
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.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Ahlquist P (1992) Bromovirus RNA replication and transcription. Curr Opin Gen Dev 2: 71–76CrossRefGoogle Scholar
  2. 2.
    Allison R, Janda M, Ahlquist P (1988) Infectious in vitro transcripts from cowpea chlorotic mottle virus cDNA clones and exchange of individual components with brome mosaic virus. J Virol 62: 3581–3588PubMedGoogle Scholar
  3. 3.
    Allison RF, Thompson C, Ahlquist P (1989) Regeneration of a functional RNA virus genome by recombination between deletion mutants and requirements for cowpea chlorotic mottle virus 3a and coat genes for systemic infection. Proc Natl Acad Sci USA 87: 1820–1824CrossRefGoogle Scholar
  4. 4.
    Broyles S, Moss B (1987) Sedimentation of an RNA polymerase complex from vaccinia virus that specifically initiates and terminates transcription. Mol Cell Biol 7: 7–14PubMedGoogle Scholar
  5. 5.
    DeJong W, Ahlquist P (1992) A hybrid plant virus made by transferring the noncapsid movement protein from a rod-shaped to an icosahedral virus is competent for systemic infection. Proc Natl Acad Sci USA 89: 6808–6812CrossRefGoogle Scholar
  6. 6.
    Felgner PL, Gadek TR, Holm M, Roman R, Chan HW, Wenz M, Northrop JP, Ringold GM, Danielsen M (1987) Lipofection: a highly efficient, lipid-mediated DNA-transfection procedure. Proc Natl Acad Sci USA 84: 7413–7417PubMedCrossRefGoogle Scholar
  7. 7.
    Francki RIB, Fauquet CM, Knudson DL, Brown F (1991) Classification and nomenclature of viruses: Fifth Report of the International Committee on Taxonomy of Viruses. Springer, Wien New York (Arch Virol [Suppl] 2 )Google Scholar
  8. 8.
    Goldbach R, LeGall O, Wellink J (1991) Alpha-like viruses in plants. Semin Virol 2: 19–25Google Scholar
  9. 9.
    Gorbalenya AE, Koonin EV (1989) Viral proteins containing the purine NTP- binding sequence pattern. Nucleic Acids Res 17: 8413–8440PubMedCrossRefGoogle Scholar
  10. 10.
    Guinea R, Carrasco L (1990) Phospholipid biosynthesis and poliovirus genome replication, two coupled phenomena. EMBO J 9:2011–2016PubMedGoogle Scholar
  11. 11.
    Haseloff J, Goelet P, Zimmern D, Ahlquist P, Dasgupta R, Kaesberg P (1984) Striking similarities in amino acid sequence among nonstructural proteins encoded by RNA viruses that have dissimilar genome organization. Proc Natl Acad Sci USA 81: 4358–4362PubMedCrossRefGoogle Scholar
  12. 12.
    Hayes RJ, Buck KW (1990) Complete replication of a eukaryotic virus RNA by a purified RNA-dependent RNA polymerase. Cell 63: 363–368PubMedCrossRefGoogle Scholar
  13. 13.
    Hodgman TC (1988) A new superfamily of replicative proteins. Nature 333: 22–23; 578 [Erratum]CrossRefGoogle Scholar
  14. 14.
    Kao C, Ahlquist P (1992) Identification of the domains required for direct interaction of the helicase-like and polymerase-like RNA replication proteins of brome mosaic virus. J Virol 66: 7293–7302PubMedGoogle Scholar
  15. 15.
    Kao C, Quadt R, Hershberger R, Ahlquist P (1992) Brome mosaic virus RNA replication proteins la and 2a form a complex in vitro. J Virol 66: 6322–6329PubMedGoogle Scholar
  16. 16.
    Kroner PA, Richards D, Traynor P, Ahlquist P (1989) Defined mutations in a small region of the brome mosaic virus 2a gene cause diverse temperature-sensitive RNA replication phenotypes. J Virol 63: 5302–5309PubMedGoogle Scholar
  17. 17.
    Kroner PA, Young B, Ahlquist P (1990) Analysis of the role of brome mosaic virus la protein domains in RNA replication, using linker insertion mutagenesis. J Virol 64: 6110–6120PubMedGoogle Scholar
  18. 18.
    Mi S, Stollar V (1991) Expression of Sindbis virus nsPl and methyltransferase activity in Escherichia coli. Virology 184: 423–427PubMedCrossRefGoogle Scholar
  19. 19.
    Molla A, Paul AV, Wimmer E (1991) Cell-free, de novo synthesis of poliovirus. Science 254: 1647–1651PubMedCrossRefGoogle Scholar
  20. 20.
    Orr JW, Newton AC (1992) Interaction of protein kinase C with phosphatidylserine. 1. Cooperativity in lipid binding. Biochemistry 31: 4661–4667Google Scholar
  21. 21.
    Perez L, Guinea R, Carrasco L (1991) Synthesis of Semliki Forest virus RNA requires continuous lipid synthesis. Virology 183: 74–82PubMedCrossRefGoogle Scholar
  22. 22.
    Quadt R, Jaspars EM J (1990) Purification and characterization of brome mosaic virus RNA-dependent RNA polymerase. Virology 178: 189–194PubMedCrossRefGoogle Scholar
  23. 23.
    Scheidel LM, Stollar V (1991) Mutations that confer resistance to mycophenolic acid and ribavirin on Sindbis virus map to the nonstructural protein nsPl. Virology 181: 490–499PubMedCrossRefGoogle Scholar
  24. 24.
    Sekimizu K, Kornberg A (1988) Cardiolipin activation of dnaA protein, the initiation protein of replication in Escherichia coli. J Biol Chem 263: 7131–7135PubMedGoogle Scholar
  25. 25.
    Traynor P, Ahlquist P (1990) Use of bromovirus RNA hybrids to map cis- and trans-acting functions in a conserved RNA replication gene. J Virol 64: 69–77PubMedGoogle Scholar
  26. 26.
    Traynor P, Young B, Ahlquist P (1991) Deletion analysis of brome mosaic virus 2a protein: effects on RNA replication and systemic spread. J Virol 65: 2807–2815PubMedGoogle Scholar
  27. 27.
    Wu SX, Ahlquist P, Kaesberg P (1992) Active complete in vitro replication of nodavirus RNA requires glycerophospholipid. Proc Natl Acad Sci USA 89: 11136–11140PubMedCrossRefGoogle Scholar
  28. 28.
    Wu SX, Kaesberg P (1991) Synthesis of template-sense, single-strand Flock house virus RNA in a cell-free replication system. Virology 183: 392–396PubMedCrossRefGoogle Scholar
  29. 29.
    Dinant M, Janda M, Kroner P, Ahlquist P (1993) Bromovirus RNA replication and transcription require compatibility between the polymerase- and helicase-like viral RNA synthesis proteins. J Virol 67 (in press)Google Scholar

Copyright information

© Springer-Verlag 1994

Authors and Affiliations

  • P. Ahlquist
    • 1
    • 2
  • S.-X. Wu
    • 1
    • 3
  • P. Kaesberg
    • 1
    • 3
  • C. C. Kao
    • 1
    • 2
  • R. Quadt
    • 1
    • 2
  • W. Dejong
    • 1
    • 2
  • R. Hershberger
    • 1
    • 2
  1. 1.Institute for Molecular VirologyUniversity of Wisconsin-MadisonMadisonUSA
  2. 2.Department of Plant PathologyUniversity of Wisconsin-MadisonMadisonUSA
  3. 3.Department of BiochemistryUniversity of Wisconsin-MadisonMadisonUSA

Personalised recommendations