1887

Abstract

RNA virus genome replication requires initiation at the precise terminus of the template RNA. To investigate the nucleotide requirements for initiation of hepatitis C virus (HCV) positive-strand RNA replication, a hammerhead ribozyme was inserted at the 5′ end of an HCV subgenomic replicon, allowing the generation of replicons with all four possible nucleotides at position 1. This analysis revealed a preference for a purine nucleotide at this position for initiation of RNA replication. The sequence requirements at positions 2–4 in the context of the J6/JFH-1 virus were also examined by selecting replication-competent virus from a pool containing randomized residues at these positions. There was strong selection for both the wild-type cytosine at position 2, and the wild-type sequence at positions 2–4 (CCU). An adenine residue was well tolerated at positions 3 and 4, which suggests that efficient RNA replication is less dependent on these residues.

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2011-05-01
2020-05-26
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References

  1. Cai Z., Liang T. J., Luo G. 2004; Effects of mutations of the initiation nucleotides on hepatitis C virus RNA replication in the cell. J Virol 78:3633–3643 [CrossRef][PubMed]
    [Google Scholar]
  2. Hughes M., Gretton S., Shelton H., Brown D. D., McCormick C. J., Angus A. G., Patel A. H., Griffin S., Harris M. 2009; A conserved proline between domains II and III of hepatitis C virus NS5A influences both RNA replication and virus assembly. J Virol 83:10788–10796 [CrossRef][PubMed]
    [Google Scholar]
  3. Kato T., Furusaka A., Miyamoto M., Date T., Yasui K., Hiramoto J., Nagayama K., Tanaka T., Wakita T. 2001; Sequence analysis of hepatitis C virus isolated from a fulminant hepatitis patient. J Med Virol 64:334–339 [CrossRef][PubMed]
    [Google Scholar]
  4. Kato T., Date T., Miyamoto M., Furusaka A., Tokushige K., Mizokami M., Wakita T. 2003; Efficient replication of the genotype 2a hepatitis C virus subgenomic replicon. Gastroenterology 125:1808–1817 [CrossRef][PubMed]
    [Google Scholar]
  5. Krieger N., Lohmann V., Bartenschlager R. 2001; Enhancement of hepatitis C virus RNA replication by cell culture-adaptive mutations. J Virol 75:4614–4624 [CrossRef][PubMed]
    [Google Scholar]
  6. Kuiken C., Yusim K., Boykin L., Richardson R. 2005; The Los Alamos hepatitis C sequence database. Bioinformatics 21:379–384 [CrossRef][PubMed]
    [Google Scholar]
  7. Lohmann V., Körner F., Koch J. O., Herian U., Theilmann L., Bartenschlager R. 1999; Replication of subgenomic hepatitis C virus RNAs in a hepatoma cell line. Science 285:110–113 [CrossRef][PubMed]
    [Google Scholar]
  8. Luo G., Hamatake R. K., Mathis D. M., Racela J., Rigat K. L., Lemm J., Colonno R. J. 2000; De novo initiation of RNA synthesis by the RNA-dependent RNA polymerase (NS5B) of hepatitis C virus. J Virol 74:851–863 [CrossRef][PubMed]
    [Google Scholar]
  9. Luo G., Xin S., Cai Z. 2003; Role of the 5′-proximal stem-loop structure of the 5′ untranslated region in replication and translation of hepatitis C virus RNA. J Virol 77:3312–3318 [CrossRef][PubMed]
    [Google Scholar]
  10. McCormick C. J., Challinor L., Macdonald A., Rowlands D. J., Harris M. 2004; Introduction of replication-competent hepatitis C virus transcripts using a tetracycline-regulable baculovirus delivery system. J Gen Virol 85:429–439 [CrossRef][PubMed]
    [Google Scholar]
  11. Pietschmann T., Kaul A., Koutsoudakis G., Shavinskaya A., Kallis S., Steinmann E., Abid K., Negro F., Dreux M. et al. 2006; Construction and characterization of infectious intragenotypic and intergenotypic hepatitis C virus chimeras. Proc Natl Acad Sci U S A 103:7408–7413 [CrossRef][PubMed]
    [Google Scholar]
  12. Shim J. H., Larson G., Wu J. Z., Hong Z. 2002; Selection of 3′-template bases and initiating nucleotides by hepatitis C virus NS5B RNA-dependent RNA polymerase. J Virol 76:7030–7039 [CrossRef][PubMed]
    [Google Scholar]
  13. Simister P., Schmitt M., Geitmann M., Wicht O., Danielson U. H., Klein R., Bressanelli S., Lohmann V. 2009; Structural and functional analysis of hepatitis C virus strain JFH1 polymerase. J Virol 83:11926–11939 [CrossRef][PubMed]
    [Google Scholar]
  14. Wakita T., Pietschmann T., Kato T., Date T., Miyamoto M., Zhao Z., Murthy K., Habermann A., Kräusslich H. G. et al. 2005; Production of infectious hepatitis C virus in tissue culture from a cloned viral genome. Nat Med 11:791–796 [CrossRef][PubMed]
    [Google Scholar]
  15. Zhong W., Uss A. S., Ferrari E., Lau J. Y., Hong Z. 2000; De novo initiation of RNA synthesis by hepatitis C virus nonstructural protein 5B polymerase. J Virol 74:2017–2022 [CrossRef][PubMed]
    [Google Scholar]
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