Abstract
Mutants of influenza Rostock virus (H7N1 subtype) were selected for resistance to amantadine hydrochloride at concentrations of the antiviral drug known to affect the function of the virus M2 transmembrane protein. Sequence analysis revealed that three mutants had no changes in M2 but contained a lysine to isoleucine substitution in the hemagglutinin (HA) membrane glycoprotein at position 58 of HA2. The mutant viruses were found to fuse membranes at a pH value 0.7 lower than wild type and to exhibit changes in the conformation of their HAs specifically at the lower pH. The homologous lysine to isoleucine substitution was introduced by site-specific mutagenesis into the HA of X-31 influenza virus (H3 subtype), which was expressed by using vaccinia virus recombinants. The expressed HA also mediated membrane fusion and changed in conformation at a pH value 0.7 lower than wild type. These results indicate that increased acid stability of the HA obviates the consequences of the inhibition of M2 function by amantadine and provide further evidence for the role of M2 in regulating the pH of vesicles involved in glycoprotein transport to the cell surface.
Full text
PDFImages in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Belshe R. B., Smith M. H., Hall C. B., Betts R., Hay A. J. Genetic basis of resistance to rimantadine emerging during treatment of influenza virus infection. J Virol. 1988 May;62(5):1508–1512. doi: 10.1128/jvi.62.5.1508-1512.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Brand C. M., Skehel J. J. Crystalline antigen from the influenza virus envelope. Nat New Biol. 1972 Aug 2;238(83):145–147. doi: 10.1038/newbio238145a0. [DOI] [PubMed] [Google Scholar]
- Bukrinskaya A. G., Vorkunova N. K., Kornilayeva G. V., Narmanbetova R. A., Vorkunova G. K. Influenza virus uncoating in infected cells and effect of rimantadine. J Gen Virol. 1982 May;60(Pt 1):49–59. doi: 10.1099/0022-1317-60-1-49. [DOI] [PubMed] [Google Scholar]
- Bukrinskaya A. G., Vorkunova N. K., Pushkarskaya N. L. Uncoating of a rimantadine-resistant variant of influenza virus in the presence of rimantadine. J Gen Virol. 1982 May;60(Pt 1):61–66. doi: 10.1099/0022-1317-60-1-61. [DOI] [PubMed] [Google Scholar]
- Daniels R. S., Downie J. C., Hay A. J., Knossow M., Skehel J. J., Wang M. L., Wiley D. C. Fusion mutants of the influenza virus hemagglutinin glycoprotein. Cell. 1985 Feb;40(2):431–439. doi: 10.1016/0092-8674(85)90157-6. [DOI] [PubMed] [Google Scholar]
- Doms R. W., Helenius A., White J. Membrane fusion activity of the influenza virus hemagglutinin. The low pH-induced conformational change. J Biol Chem. 1985 Mar 10;260(5):2973–2981. [PubMed] [Google Scholar]
- Hay A. J., Wolstenholme A. J., Skehel J. J., Smith M. H. The molecular basis of the specific anti-influenza action of amantadine. EMBO J. 1985 Nov;4(11):3021–3024. doi: 10.1002/j.1460-2075.1985.tb04038.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hayden F. G., Belshe R. B., Clover R. D., Hay A. J., Oakes M. G., Soo W. Emergence and apparent transmission of rimantadine-resistant influenza A virus in families. N Engl J Med. 1989 Dec 21;321(25):1696–1702. doi: 10.1056/NEJM198912213212502. [DOI] [PubMed] [Google Scholar]
- Kawaoka Y., Yamnikova S., Chambers T. M., Lvov D. K., Webster R. G. Molecular characterization of a new hemagglutinin, subtype H14, of influenza A virus. Virology. 1990 Dec;179(2):759–767. doi: 10.1016/0042-6822(90)90143-f. [DOI] [PubMed] [Google Scholar]
- Kunkel T. A., Roberts J. D., Zakour R. A. Rapid and efficient site-specific mutagenesis without phenotypic selection. Methods Enzymol. 1987;154:367–382. doi: 10.1016/0076-6879(87)54085-x. [DOI] [PubMed] [Google Scholar]
- Lamb R. A., Zebedee S. L., Richardson C. D. Influenza virus M2 protein is an integral membrane protein expressed on the infected-cell surface. Cell. 1985 Mar;40(3):627–633. doi: 10.1016/0092-8674(85)90211-9. [DOI] [PubMed] [Google Scholar]
- Mackett M., Smith G. L., Moss B. General method for production and selection of infectious vaccinia virus recombinants expressing foreign genes. J Virol. 1984 Mar;49(3):857–864. doi: 10.1128/jvi.49.3.857-864.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Marsh M., Helenius A. Virus entry into animal cells. Adv Virus Res. 1989;36:107–151. doi: 10.1016/S0065-3527(08)60583-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Martin K., Helenius A. Nuclear transport of influenza virus ribonucleoproteins: the viral matrix protein (M1) promotes export and inhibits import. Cell. 1991 Oct 4;67(1):117–130. doi: 10.1016/0092-8674(91)90576-k. [DOI] [PubMed] [Google Scholar]
- Nobusawa E., Aoyama T., Kato H., Suzuki Y., Tateno Y., Nakajima K. Comparison of complete amino acid sequences and receptor-binding properties among 13 serotypes of hemagglutinins of influenza A viruses. Virology. 1991 Jun;182(2):475–485. doi: 10.1016/0042-6822(91)90588-3. [DOI] [PubMed] [Google Scholar]
- Patel D. D., Ray C. A., Drucker R. P., Pickup D. J. A poxvirus-derived vector that directs high levels of expression of cloned genes in mammalian cells. Proc Natl Acad Sci U S A. 1988 Dec;85(24):9431–9435. doi: 10.1073/pnas.85.24.9431. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ruigrok R. W., Hirst E. M., Hay A. J. The specific inhibition of influenza A virus maturation by amantadine: an electron microscopic examination. J Gen Virol. 1991 Jan;72(Pt 1):191–194. doi: 10.1099/0022-1317-72-1-191. [DOI] [PubMed] [Google Scholar]
- Skehel J. J., Bayley P. M., Brown E. B., Martin S. R., Waterfield M. D., White J. M., Wilson I. A., Wiley D. C. Changes in the conformation of influenza virus hemagglutinin at the pH optimum of virus-mediated membrane fusion. Proc Natl Acad Sci U S A. 1982 Feb;79(4):968–972. doi: 10.1073/pnas.79.4.968. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Stegmann T., Doms R. W., Helenius A. Protein-mediated membrane fusion. Annu Rev Biophys Biophys Chem. 1989;18:187–211. doi: 10.1146/annurev.bb.18.060189.001155. [DOI] [PubMed] [Google Scholar]
- Steinhauer D. A., Wharton S. A., Wiley D. C., Skehel J. J. Deacylation of the hemagglutinin of influenza A/Aichi/2/68 has no effect on membrane fusion properties. Virology. 1991 Sep;184(1):445–448. doi: 10.1016/0042-6822(91)90867-b. [DOI] [PubMed] [Google Scholar]
- Sugrue R. J., Bahadur G., Zambon M. C., Hall-Smith M., Douglas A. R., Hay A. J. Specific structural alteration of the influenza haemagglutinin by amantadine. EMBO J. 1990 Nov;9(11):3469–3476. doi: 10.1002/j.1460-2075.1990.tb07555.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sugrue R. J., Hay A. J. Structural characteristics of the M2 protein of influenza A viruses: evidence that it forms a tetrameric channel. Virology. 1991 Feb;180(2):617–624. doi: 10.1016/0042-6822(91)90075-M. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Venkatesan S., Baroudy B. M., Moss B. Distinctive nucleotide sequences adjacent to multiple initiation and termination sites of an early vaccinia virus gene. Cell. 1981 Sep;25(3):805–813. doi: 10.1016/0092-8674(81)90188-4. [DOI] [PubMed] [Google Scholar]
- Weis W. I., Brünger A. T., Skehel J. J., Wiley D. C. Refinement of the influenza virus hemagglutinin by simulated annealing. J Mol Biol. 1990 Apr 20;212(4):737–761. doi: 10.1016/0022-2836(90)90234-D. [DOI] [PubMed] [Google Scholar]
- Weis W. I., Cusack S. C., Brown J. H., Daniels R. S., Skehel J. J., Wiley D. C. The structure of a membrane fusion mutant of the influenza virus haemagglutinin. EMBO J. 1990 Jan;9(1):17–24. doi: 10.1002/j.1460-2075.1990.tb08075.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wharton S. A., Skehel J. J., Wiley D. C. Studies of influenza haemagglutinin-mediated membrane fusion. Virology. 1986 Feb;149(1):27–35. doi: 10.1016/0042-6822(86)90083-8. [DOI] [PubMed] [Google Scholar]
- White J. M. Viral and cellular membrane fusion proteins. Annu Rev Physiol. 1990;52:675–697. doi: 10.1146/annurev.ph.52.030190.003331. [DOI] [PubMed] [Google Scholar]
- White J. M., Wilson I. A. Anti-peptide antibodies detect steps in a protein conformational change: low-pH activation of the influenza virus hemagglutinin. J Cell Biol. 1987 Dec;105(6 Pt 2):2887–2896. doi: 10.1083/jcb.105.6.2887. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wiley D. C., Skehel J. J. The structure and function of the hemagglutinin membrane glycoprotein of influenza virus. Annu Rev Biochem. 1987;56:365–394. doi: 10.1146/annurev.bi.56.070187.002053. [DOI] [PubMed] [Google Scholar]
- Wilson I. A., Skehel J. J., Wiley D. C. Structure of the haemagglutinin membrane glycoprotein of influenza virus at 3 A resolution. Nature. 1981 Jan 29;289(5796):366–373. doi: 10.1038/289366a0. [DOI] [PubMed] [Google Scholar]
- Zebedee S. L., Lamb R. A. Influenza A virus M2 protein: monoclonal antibody restriction of virus growth and detection of M2 in virions. J Virol. 1988 Aug;62(8):2762–2772. doi: 10.1128/jvi.62.8.2762-2772.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Zhirnov O. P. Solubilization of matrix protein M1/M from virions occurs at different pH for orthomyxo- and paramyxoviruses. Virology. 1990 May;176(1):274–279. doi: 10.1016/0042-6822(90)90253-n. [DOI] [PubMed] [Google Scholar]