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PMID: 17936324 Published · ppublish English Journal Article Research Support, N.I.H., Extramural

Analysis of residues near the fusion peptide in the influenza hemagglutinin structure for roles in triggering membrane fusion.

Virology ·Vol. 370 ·No. 2 ·2008-01-20 ·Pages 403-14

Thoennes S, Li ZN, Lee BJ, Langley WA, Skehel JJ, Russell RJ, Steinhauer DA

Abstract

Influenza virus entry occurs in endosomes, where acidification triggers irreversible conformational changes of the hemagglutinin glycoprotein (HA) that are required for membrane fusion. The acid-induced HA structural rearrangements have been well documented, and several models have been proposed to relate these to the process of membrane fusion. However, details regarding the role of specific residues in the initiation of structural rearrangements and membrane fusion are lacking. Here we report the results of studies on the HA of A/Aichi/2/68 virus (H3 subtype), in which mutants with changes at several ionizable residues in the vicinity of the "fusion peptide" were analyzed for their effects on the pH at which conformational changes and membrane fusion occur. A variety of phenotypes was obtained, including examples of substitutions that lead to an increase in HA stability at reduced pH. Of particular note was the observation that a histidine to tyrosine substitution at HA1 position 17 resulted in a decrease in pH at which HA structural changes and membrane fusion take place by 0.3 relative to WT. The results are discussed in relation to possible mechanisms by which HA structural rearrangements are initiated at low pH and clade-specific differences near the fusion peptide.

MeSH Terms
Amino Acid Substitution Animals Cell Line Cricetinae Hemagglutinin Glycoproteins, Influenza Virus/chemistry,genetics,physiology Humans Hydrogen-Ion Concentration Influenza A Virus, H3N2 Subtype/genetics,physiology Membrane Fusion/genetics,physiology Models, Molecular Mutagenesis, Site-Directed Phylogeny Protein Conformation Recombinant Proteins/chemistry,genetics,metabolism Trypsin Viral Fusion Proteins/chemistry,genetics,physiology
Chemicals
Hemagglutinin Glycoproteins, Influenza Virus Recombinant Proteins Viral Fusion Proteins Trypsin
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Thoennes Sudha
Department of Microbiology and Immunology, Emory University School of Medicine, 1510 Clifton Road, Atlanta, GA 30322, USA.
Li Zhu-Nan
Lee Byeong-Jae
Langley William A
Skehel John J
Russell Rupert J
Steinhauer David A
References (59)
59 references, click to expand
  1. Viral entry.
    Curr Top Microbiol Immunol. 2005;285:1-23 PMID: 15609499
  2. Studies of the membrane fusion activities of fusion peptide mutants of influenza virus hemagglutinin.
    J Virol. 1995 Nov;69(11):6643-51 PMID: 7474073
  3. Inhibition of the fusion-inducing conformational change of influenza hemagglutinin by benzoquinones and hydroquinones.
    Biochemistry. 1993 Mar 30;32(12):2967-78 PMID: 8457561
  4. Murine leukemia virus R Peptide inhibits influenza virus hemagglutinin-induced membrane fusion.
    J Virol. 2006 Jun;80(12):6106-14 PMID: 16731949
  5. The structure and function of the hemagglutinin membrane glycoprotein of influenza virus.
    Annu Rev Biochem. 1987;56:365-94 PMID: 3304138
  6. Electron microscopy of influenza haemagglutinin-monoclonal antibody complexes.
    Virology. 1983 Dec;131(2):308-14 PMID: 6689229
  7. Structural studies on viral escape from antibody neutralization.
    Curr Top Microbiol Immunol. 2001;260:55-64 PMID: 11443881
  8. How viruses enter animal cells.
    Science. 2004 Apr 9;304(5668):237-42 PMID: 15073366
  9. Paramyxovirus membrane fusion: lessons from the F and HN atomic structures.
    Virology. 2006 Jan 5;344(1):30-7 PMID: 16364733
  10. Studies on the mechanism of membrane fusion: site-specific mutagenesis of the hemagglutinin of influenza virus.
    J Cell Biol. 1986 Jan;102(1):11-23 PMID: 3753607
  11. Salicylamide inhibitors of influenza virus fusion.
    Bioorg Med Chem Lett. 2000 Aug 7;10(15):1649-52 PMID: 10937716
  12. pH-dependent changes in photoaffinity labeling patterns of the H1 influenza virus hemagglutinin by using an inhibitor of viral fusion.
    J Virol. 1999 Mar;73(3):1785-94 PMID: 9971755
  13. Studies on the adaptation of influenza viruses to MDCK cells.
    EMBO J. 1984 Dec 20;3(13):3329-32 PMID: 6526017
  14. Studies of influenza haemagglutinin-mediated membrane fusion.
    Virology. 1986 Feb;149(1):27-35 PMID: 3946080
  15. Structure of the haemagglutinin membrane glycoprotein of influenza virus at 3 A resolution.
    Nature. 1981 Jan 29;289(5796):366-73 PMID: 7464906
  16. Characterization of a novel influenza A virus hemagglutinin subtype (H16) obtained from black-headed gulls.
    J Virol. 2005 Mar;79(5):2814-22 PMID: 15709000
  17. Structure of the hemagglutinin precursor cleavage site, a determinant of influenza pathogenicity and the origin of the labile conformation.
    Cell. 1998 Oct 30;95(3):409-17 PMID: 9814710
  18. Introduction of intersubunit disulfide bonds in the membrane-distal region of the influenza hemagglutinin abolishes membrane fusion activity.
    Cell. 1992 Feb 21;68(4):635-45 PMID: 1739972
  19. 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):2887-96 PMID: 2447101
  20. Amantadine selection of a mutant influenza virus containing an acid-stable hemagglutinin glycoprotein: evidence for virus-specific regulation of the pH of glycoprotein transport vesicles.
    Proc Natl Acad Sci U S A. 1991 Dec 15;88(24):11525-9 PMID: 1763066
  21. Deacylation of the hemagglutinin of influenza A/Aichi/2/68 has no effect on membrane fusion properties.
    Virology. 1991 Sep;184(1):445-8 PMID: 1871979
  22. Structure of influenza haemagglutinin at neutral and at fusogenic pH by electron cryo-microscopy.
    FEBS Lett. 1999 Dec 17;463(3):255-9 PMID: 10606732
  23. Studies using double mutants of the conformational transitions in influenza hemagglutinin required for its membrane fusion activity.
    Proc Natl Acad Sci U S A. 1996 Nov 12;93(23):12873-8 PMID: 8917512
  24. Enhancement of the infectivity of influenza A and B viruses by proteolytic cleavage of the hemagglutinin polypeptide.
    Virology. 1975 Dec;68(2):440-54 PMID: 128196
  25. H1 and H7 influenza haemagglutinin structures extend a structural classification of haemagglutinin subtypes.
    Virology. 2004 Aug 1;325(2):287-96 PMID: 15246268
  26. The structure and receptor binding properties of the 1918 influenza hemagglutinin.
    Science. 2004 Mar 19;303(5665):1838-42 PMID: 14764886
  27. Activation of influenza A viruses by trypsin treatment.
    Virology. 1975 Dec;68(2):426-39 PMID: 173078
  28. Conformational changes and fusion activity of influenza virus hemagglutinin of the H2 and H3 subtypes: effects of acid pretreatment.
    J Virol. 1990 Aug;64(8):3824-32 PMID: 2196382
  29. Unwelcome guests with master keys: how HIV enters cells and how it can be stopped.
    Top HIV Med. 2004 Oct-Nov;12(4):100-3 PMID: 15516706
  30. Structure of influenza haemagglutinin at the pH of membrane fusion.
    Nature. 1994 Sep 1;371(6492):37-43 PMID: 8072525
  31. Acid-induced changes in thermal stability and fusion activity of influenza hemagglutinin.
    Biochemistry. 2002 Feb 12;41(6):2044-54 PMID: 11827552
  32. Analyses of the antigenicity of influenza haemagglutinin at the pH optimum for virus-mediated membrane fusion.
    J Gen Virol. 1983 Aug;64 (Pt 8):1657-62 PMID: 6192202
  33. Mechanism of membrane fusion by viral envelope proteins.
    Adv Virus Res. 2005;64:231-61 PMID: 16139596
  34. Molecular characterization of a new hemagglutinin, subtype H14, of influenza A virus.
    Virology. 1990 Dec;179(2):759-67 PMID: 2238469
  35. Selection of recombinant vaccinia viruses on the basis of plaque formation.
    Gene. 1995 Jun 9;158(2):157-62 PMID: 7607536
  36. The many mechanisms of viral membrane fusion proteins.
    Curr Top Microbiol Immunol. 2005;285:25-66 PMID: 15609500
  37. Virus membrane fusion proteins: biological machines that undergo a metamorphosis.
    Biosci Rep. 2000 Dec;20(6):597-612 PMID: 11426696
  38. Class II virus membrane fusion proteins.
    Virology. 2006 Jan 5;344(1):38-47 PMID: 16364734
  39. Receptor binding and membrane fusion in virus entry: the influenza hemagglutinin.
    Annu Rev Biochem. 2000;69:531-69 PMID: 10966468
  40. Characterization of a novel influenza hemagglutinin, H15: criteria for determination of influenza A subtypes.
    Virology. 1996 Mar 15;217(2):508-16 PMID: 8610442
  41. The structural biology of type I viral membrane fusion.
    Nat Rev Mol Cell Biol. 2003 Apr;4(4):309-19 PMID: 12671653
  42. Plaque formation by influenza viruses in the presence of trypsin.
    J Gen Virol. 1974 Dec;25(3):351-7 PMID: 4475092
  43. Studies on influenza haemagglutinin fusion peptide mutants generated by reverse genetics.
    EMBO J. 2001 Aug 15;20(16):4432-42 PMID: 11500371
  44. Variant influenza virus hemagglutinin that induces fusion at elevated pH.
    J Virol. 1986 Feb;57(2):603-13 PMID: 3003392
  45. Structure of influenza virus haemagglutinin complexed with a neutralizing antibody.
    Nature. 1995 Jul 6;376(6535):92-4 PMID: 7596443
  46. Transient changes of the conformation of hemagglutinin of influenza virus at low pH detected by time-resolved circular dichroism spectroscopy.
    J Biol Chem. 1997 Apr 11;272(15):9764-70 PMID: 9092509
  47. Novel quinolizidine salicylamide influenza fusion inhibitors.
    Bioorg Med Chem Lett. 1999 Aug 2;9(15):2177-80 PMID: 10465540
  48. Intermonomer disulfide bonds impair the fusion activity of influenza virus hemagglutinin.
    J Virol. 1992 Aug;66(8):4940-50 PMID: 1629960
  49. Fusion mutants of the influenza virus hemagglutinin glycoprotein.
    Cell. 1985 Feb;40(2):431-9 PMID: 3967299
  50. Mechanisms of viral membrane fusion and its inhibition.
    Annu Rev Biochem. 2001;70:777-810 PMID: 11395423
  51. Adaptation of egg-grown and transfectant influenza viruses for growth in mammalian cells: selection of hemagglutinin mutants with elevated pH of membrane fusion.
    Virology. 1997 Jul 7;233(2):402-10 PMID: 9217063
  52. Avian sarcoma and leukosis virus-receptor interactions: from classical genetics to novel insights into virus-cell membrane fusion.
    Virology. 2006 Jan 5;344(1):25-9 PMID: 16364732
  53. 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):968-72 PMID: 6951181
  54. Early steps of the conformational change of influenza virus hemagglutinin to a fusion active state: stability and energetics of the hemagglutinin.
    Biochim Biophys Acta. 2003 Jul 11;1614(1):3-13 PMID: 12873761
  55. H5 avian and H9 swine influenza virus haemagglutinin structures: possible origin of influenza subtypes.
    EMBO J. 2002 Mar 1;21(5):865-75 PMID: 11867515
  56. Conformational intermediates and fusion activity of influenza virus hemagglutinin.
    J Virol. 1999 Jun;73(6):4567-74 PMID: 10233915
  57. Molecular mechanism underlying the action of a novel fusion inhibitor of influenza A virus.
    J Virol. 1997 May;71(5):4062-70 PMID: 9094684
  58. Comparison of complete amino acid sequences and receptor-binding properties among 13 serotypes of hemagglutinins of influenza A viruses.
    Virology. 1991 Jun;182(2):475-85 PMID: 2024485
  59. Structure-based identification of an inducer of the low-pH conformational change in the influenza virus hemagglutinin: irreversible inhibition of infectivity.
    J Virol. 1997 Nov;71(11):8808-20 PMID: 9343241
Article Info
Journal
Virology
Abbr.
Virology
ISSN
0042-6822
DOI
10.1016/j.virol.2007.08.035
Published
2008-01-20
Epub
2007-00-23
Pages
403-14
Language
English
Region
United States
NLM ID
0110674
PMCID
PMC2212604
Subset
IM
Grants
NIGMS NIH HHS · R01 GM066870-05 · United States
NIAID NIH HHS · R21 AI053359-01A1 · United States
NIAID NIH HHS · AI/EB53359 · United States
NIAID NIH HHS · HHSN266200700006C · United States
Medical Research Council · MC_U117512711 · United Kingdom
NIAID NIH HHS · R13 AI066870 · United States
NIGMS NIH HHS · R01 GM066870-03 · United States
NIAID NIH HHS · AI66870 · United States
NIGMS NIH HHS · R01 GM066870-02 · United States
NIGMS NIH HHS · R01 GM066870-04 · United States
NIGMS NIH HHS · R01 GM066870-01 · United States
NIAID NIH HHS · R21 AI053359-02 · United States
External Links
PubMed source DOI: 10.1016/j.virol.2007.08.035 PMC Full Text View journal details
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