1887

Abstract

The genetic stability of attenuated (HIV-1) variants harbouring mutations (Gly or Lys) of Asn17, the protease-cleavage site of the proximal zinc finger of the nucleocapsid protein, was studied. All possible codons for the Gly mutants were tested as starting sequences. Long-term replication assays revealed that the mutants were unstable; mutations of Gly17 to Arg, Ala, Ser and Cys, as well as a Lys17Asn reversion, were observed. Replication kinetic assays in H9 cells revealed that the replication of Ala, Ser and Arg mutants was improved substantially compared with the Gly variant; the infectivity of Ala17 and Ser17 viruses was equal to, and that of Arg17 was almost equal to, the infectivity of the wild-type virus. Kinetic analysis of the cleavage of oligopeptides representing the corresponding nucleocapsid-cleavage sites revealed that all mutations improved cleavability, in good agreement with the previously proposed role of nucleocapsid cleavage in HIV-1 replication.

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2006-04-01
2019-11-22
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References

  1. Amarasinghe, G. K., De Guzman, R. N., Turner, R. B., Chancellor, K. J., Wu, Z. R. & Summers, M. F. ( 2000; ). NMR structure of the HIV-1 nucleocapsid protein bound to stem–loop SL2 of the Ψ-RNA packaging signal. Implications for genome recognition. J Mol Biol 301, 491–511.[CrossRef]
    [Google Scholar]
  2. Baboonian, C., Dalgleish, A., Bountiff, L., Gross, J., Oroszlan, S., Rickett, G., Smith-Burchnell, C., Troke, P. & Merson, J. ( 1991; ). HIV-1 proteinase is required for synthesis of pro-viral DNA. Biochem Biophys Res Commun 179, 17–24.[CrossRef]
    [Google Scholar]
  3. Bampi, C., Jacquenet, S., Lener, D., Décimo, D. & Darlix, J.-L. ( 2004; ). The chaperoning and assistance roles of the HIV-1 nucleocapsid protein in proviral DNA synthesis and maintenance. Int J Biochem Cell Biol 36, 1668–1686.[CrossRef]
    [Google Scholar]
  4. Coffin, J. M. ( 1995; ). HIV population dynamics in vivo: implications for genetic variation, pathogenesis, and therapy. Science 267, 483–489.[CrossRef]
    [Google Scholar]
  5. De Guzman, R. N., Wu, Z. R., Stalling, C. C., Pappalardo, L., Borer, P. N. & Summers, M. F. ( 1998; ). Structure of the HIV-1 nucleocapsid protein bound to the SL3 Ψ-RNA recognition element. Science 279, 384–388.[CrossRef]
    [Google Scholar]
  6. Dorfman, T., Luban, J., Goff, S. P., Haseltine, W. A. & Göttlinger, H. G. ( 1993; ). Mapping of functionally important residues of a cysteine-histidine box in the human immunodeficiency virus type 1 nucleocapsid protein. J Virol 67, 6159–6169.
    [Google Scholar]
  7. Goto, T., Nakano, T., Kohno, T., Morimatsu, S., Morita, C., Hong, W., Kiso, Y., Nakai, M. & Sano, K. ( 2001; ). Targets of a protease inhibitor, KNI-272, in HIV-1-infected cells. J Med Virol 63, 203–209.[CrossRef]
    [Google Scholar]
  8. Hsu, M. & Wainberg, M. A. ( 2000; ). Interactions between human immunodeficiency virus type 1 reverse transcriptase, tRNA primer, and nucleocapsid protein during reverse transcription. J Hum Virol 3, 16–26.
    [Google Scholar]
  9. Huang, M., Maynard, A., Turpin, J. A., Graham, L., Janini, G. M., Covell, D. G. & Rice, W. G. ( 1998; ). Anti-HIV agents that selectively target retroviral nucleocapsid protein zinc fingers without affecting cellular zinc finger proteins. J Med Chem 41, 1371–1381.[CrossRef]
    [Google Scholar]
  10. Jacobsen, H., Ahlborn-Laake, L., Gugel, R. & Mous, J. ( 1992; ). Progression of early steps of human immunodeficiency virus type 1 replication in the presence of an inhibitor of viral protease. J Virol 66, 5087–5091.
    [Google Scholar]
  11. Kaplan, A. H., Manchester, M., Smith, T., Yang, Y. L. & Swanstrom, R. ( 1996; ). Conditional human immunodeficiency virus type 1 protease mutants show no role for the viral protease early in virus replication. J Virol 70, 5840–5844.
    [Google Scholar]
  12. Martínez, M. A., Sala, M., Vartanian, J.-P. & Wain-Hobson, S. ( 1995; ). Reverse transcriptase and substrate dependence of the RNA hypermutagenesis reaction. Nucleic Acids Res 23, 2573–2578.[CrossRef]
    [Google Scholar]
  13. Musah, R. A. ( 2004; ). The HIV-1 nucleocapsid zinc finger protein as a target of antiretroviral therapy. Curr Top Med Chem 4, 1605–1622.[CrossRef]
    [Google Scholar]
  14. Nagy, K., Young, M., Baboonian, C., Merson, J., Whittle, P. & Oroszlan, S. ( 1994; ). Antiviral activity of human immunodeficiency virus type 1 protease inhibitors in a single cycle of infection: evidence for a role of protease in the early phase. J Virol 68, 757–765.
    [Google Scholar]
  15. Panther, L. A., Coombs, R. W., Aung, S. A., dela Rosa, C., Gretch, D. & Corey, L. ( 1999; ). Unintegrated HIV-1 circular 2-LTR proviral DNA as a marker of recently infected cells: relative effect of recombinant CD4, zidovudine, and saquinavir in vitro. J Med Virol 58, 165–173.[CrossRef]
    [Google Scholar]
  16. Popovic, M., Sarngadharan, M. G., Read, E. & Gallo, R. C. ( 1984; ). Detection, isolation, and continuous production of cytopathic retroviruses (HTLV-III) from patients with AIDS and pre-AIDS. Science 224, 497–500.[CrossRef]
    [Google Scholar]
  17. Rein, A., Henderson, L. E. & Levin, J. G. ( 1998; ). Nucleic-acid-chaperone activity of retroviral nucleocapsid proteins: significance for viral replication. Trends Biochem Sci 23, 297–301.[CrossRef]
    [Google Scholar]
  18. Roberts, M. M. & Oroszlan, S. ( 1989; ). The preparation and biochemical characterization of intact capsids of equine infectious anemia virus. Biochem Biophys Res Commun 160, 486–494.[CrossRef]
    [Google Scholar]
  19. Roberts, M. M. & Oroszlan, S. ( 1990; ). The action of retroviral protease in various phases of virus replication. In Retroviral Proteases: Control of Maturation and Morphogenesis, pp. 131–139. Edited by L. H. Pearl. London: Macmillan Press.
  20. Roberts, M. M., Copeland, T. D. & Oroszlan, S. ( 1991a; ). In situ processing of a retroviral nucleocapsid protein by the viral proteinase. Protein Eng 4, 695–700.[CrossRef]
    [Google Scholar]
  21. Roberts, M. M., Volker, E., Copeland, T. D., Nagashima, K., Cassell, M. B., Briggs, C. J. & Oroszlan, S. ( 1991b; ). Regulated proteolytic processing within mature retroviral capsids. In Advances in Molecular Biology and Targeted Treatment for AIDS, pp. 273–280. Edited by A. Kumar. New York: Plenum.
  22. Tözsér, J. & Oroszlan, S. ( 2003; ). Proteolytic events of HIV-1 replication as targets for therapeutic intervention. Curr Pharm Des 9, 1803–1815.[CrossRef]
    [Google Scholar]
  23. Tözsér, J., Friedman, D., Weber, I. T., Blaha, I. & Oroszlan, S. ( 1993; ). Studies on the substrate specificity of the proteinase of equine infectious anemia virus using oligopeptide substrates. Biochemistry 32, 3347–3353.[CrossRef]
    [Google Scholar]
  24. Tözsér, J., Shulenin, S., Louis, J. M., Copeland, T. D. & Oroszlan, S. ( 2004; ). In vitro processing of HIV-1 nucleocapsid protein by the viral proteinase: effects of amino acid substitutions at the scissile bond in the proximal zinc finger sequence. Biochemistry 43, 4304–4312.[CrossRef]
    [Google Scholar]
  25. Uchida, H., Maeda, Y. & Mitsuya, H. ( 1997; ). HIV-1 protease does not play a critical role in the early stages of HIV-1 infection. Antiviral Res 36, 107–113.[CrossRef]
    [Google Scholar]
  26. Vartanian, J.-P., Meyerhans, A., Åsjö, B. & Wain-Hobson, S. ( 1991; ). Selection, recombination, and G→A hypermutation of human immunodeficiency virus type 1 genomes. J Virol 65, 1779–1788.
    [Google Scholar]
  27. Venaud, S., Yahi, N., Fehrentz, J. L., Guettari, N., Nisato, D., Hirsch, I. & Chermann, J. C. ( 1992; ). Inhibition of HIV by an anti-HIV protease synthetic peptide blocks an early step of viral replication. Res Virol 143, 311–319.[CrossRef]
    [Google Scholar]
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