Structure of the Hemagglutinin Precursor Cleavage Site, a Determinant of Influenza Pathogenicity and the Origin of the Labile Conformation

The membrane fusion potential of influenza HA, like many viral membrane-fusion glycoproteins, is generated by proteolytic cleavage of a biosynthetic precursor. The three-dimensional structure of ectodomain of the precursor HA0 has been determined and compared with that of cleaved HA. The cleavage site is a prominent surface loop adjacent to a novel cavity; cleavage results in structural rearrangements in which the nonpolar amino acids near the new amino terminus bury ionizable residues in the cavity that are implicated in the low-pH-induced conformational change. Amino acid insertions at the cleavage site in HAs of virulent avian viruses and those of viruses isolated from the recent severe outbreak of influenza in humans in Hong Kong would extend this surface loop, facilitating intracellular cleavage.

[1]  C. Naeve,et al.  Is virulence of H5N2 influenza viruses in chickens associated with loss of carbohydrate from the hemagglutinin? , 1984, Virology.

[2]  J. Skehel,et al.  Studies of the membrane fusion activities of fusion peptide mutants of influenza virus hemagglutinin , 1995, Journal of virology.

[3]  R. Webster,et al.  Influenza virus a pathogenicity: The pivotal role of hemagglutinin , 1987, Cell.

[4]  J. Skehel,et al.  Structure of influenza haemagglutinin at the pH of membrane fusion , 1994, Nature.

[5]  W E Stites,et al.  In a staphylococcal nuclease mutant the side-chain of a lysine replacing valine 66 is fully buried in the hydrophobic core. , 1991, Journal of molecular biology.

[6]  T. Matthews,et al.  Peptides corresponding to a predictive alpha-helical domain of human immunodeficiency virus type 1 gp41 are potent inhibitors of virus infection. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[7]  J. Skehel,et al.  The structure and function of the hemagglutinin membrane glycoprotein of influenza virus. , 1987, Annual review of biochemistry.

[8]  Experimental measurement of the J-integral , 1984 .

[9]  J. Skehel,et al.  Studies on the structure of the influenza virus haemagglutinin at the pH of membrane fusion. , 1988, The Journal of general virology.

[10]  C. Ward,et al.  Structure of the influenza virus hemagglutinin. , 1981, Current topics in microbiology and immunology.

[11]  E. Lattman,et al.  Experimental measurement of the effective dielectric in the hydrophobic core of a protein. , 1997, Biophysical chemistry.

[12]  P S Kim,et al.  Influenza hemagglutinin is spring-loaded by a metastable native conformation. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[13]  Collaborative Computational,et al.  The CCP4 suite: programs for protein crystallography. , 1994, Acta crystallographica. Section D, Biological crystallography.

[14]  N. Cox,et al.  Characterization of an avian influenza A (H5N1) virus isolated from a child with a fatal respiratory illness. , 1998, Science.

[15]  R. Webster,et al.  Interplay between carbohydrate in the stalk and the length of the connecting peptide determines the cleavability of influenza virus hemagglutinin , 1989, Journal of virology.

[16]  R. Rott,et al.  Nonhomologous recombination between the hemagglutinin gene and the nucleoprotein gene of an influenza virus. , 1994, Virology.

[17]  Benedikt Westermann,et al.  SNAREpins: Minimal Machinery for Membrane Fusion , 1998, Cell.

[18]  Q. Sattentau,et al.  Conformational changes induced in the human immunodeficiency virus envelope glycoprotein by soluble CD4 binding , 1991, The Journal of experimental medicine.

[19]  R. Webster,et al.  Glycosylation affects cleavage of an H5N2 influenza virus hemagglutinin and regulates virulence. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[20]  R. Rott,et al.  Increased viral pathogenicity after insertion of a 28S ribosomal RNA sequence into the haemagglutinin gene of an influenza virus , 1989, Nature.

[21]  P. Kraulis.  A program to produce both detailed and schematic plots of protein structures , 1991 .

[22]  S. Cusack,et al.  Structure of the influenza virus haemagglutinin complexed with its receptor, sialic acid , 1988, Nature.

[23]  I. Wilson,et al.  Structure of the haemagglutinin membrane glycoprotein of influenza virus at 3 Å resolution , 1981, Nature.

[24]  S. Watowich,et al.  Crystal structures of influenza virus hemagglutinin in complex with high-affinity receptor analogs. , 1994, Structure.

[25]  S. Ohnishi,et al.  Activation of influenza virus by acidic media causes hemolysis and fusion of erythrocytes , 1980, FEBS letters.

[26]  B. Matthews,et al.  Structural and thermodynamic consequences of burying a charged residue within the hydrophobic core of T4 lysozyme. , 1991, Biochemistry.

[27]  D. M. Ryan,et al.  Rational design of potent sialidase-based inhibitors of influenza virus replication , 1993, Nature.

[28]  A. Hay.  The Mechanism of Action of Amantadine and Rimantadine Against Influenza Viruses , 1989 .

[29]  H. Klenk,et al.  Activation of influenza A viruses by trypsin treatment. , 1975, Virology.

[30]  Reinhard Jahn,et al.  Structure and Conformational Changes in NSF and Its Membrane Receptor Complexes Visualized by Quick-Freeze/Deep-Etch Electron Microscopy , 1997, Cell.

[31]  H. Klenk,et al.  Host cell proteases controlling virus pathogenicity. , 1994, Trends in microbiology.

[32]  Maricarmen García,et al.  Virulence-associated sequence duplication at the hemagglutinin cleavage site of avian influenza viruses. , 1997, Virus research.

[33]  H. Klenk,et al.  Hemagglutinin activation of pathogenic avian influenza viruses of serotype H7 requires the protease recognition motif R-X-K/R-R☆ , 1992, Virology.

[34]  J. Henneberry,et al.  Chapter 11 Fusion Activity of the Hemagglutinin of Influenza Virus , 1988 .

[35]  Deborah Fass,et al.  Core Structure of gp41 from the HIV Envelope Glycoprotein , 1997, Cell.

[36]  W. Weissenhorn,et al.  A soluble domain of the membrane-anchoring chain of influenza virus hemagglutinin (HA2) folds in Escherichia coli into the low-pH-induced conformation. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[37]  J. Skehel,et al.  Introduction of intersubunit disulfide bonds in the membrane-distal region of the influenza hemagglutinin abolishes membrane fusion activity , 1992, Cell.

[38]  N. Katunuma,et al.  Isolation and characterization of a novel trypsin-like protease found in rat bronchiolar epithelial Clara cells. A possible activator of the viral fusion glycoprotein. , 1992, The Journal of biological chemistry.

[39]  The refinement of the haemagglutinin membrane glycoprotein of influenza virus , 1986 .

[40]  M. Vey,et al.  Influenza virus hemagglutinin with multibasic cleavage site is activated by furin, a subtilisin‐like endoprotease. , 1992, The EMBO journal.

[41]  H. Klenk,et al.  Mutations at the cleavage site of the hemagglutinin after the pathogenicity of influenza virus A/chick/Penn/83 (H5N2). , 1989, Virology.

[42]  Jorge Navaza,et al.  [33] AMoRe: An automated molecular replacement program package. , 1997, Methods in enzymology.

[43]  S. Harrison,et al.  Atomic structure of the ectodomain from HIV-1 gp41 , 1997, Nature.

[44]  D. Bastia,et al.  Rapid purification of a cloned gene product by genetic fusion and site-specific proteolysis. , 1984, Proceedings of the National Academy of Sciences of the United States of America.

[45]  I D Kuntz,et al.  Structure-based identification of an inducer of the low-pH conformational change in the influenza virus hemagglutinin: irreversible inhibition of infectivity , 1997, Journal of virology.

[46]  P. Choppin,et al.  Enhancement of the infectivity of influenza A and B viruses by proteolytic cleavage of the hemagglutinin polypeptide. , 1975, Virology.

[47]  D. Wiley,et al.  Fusion mutants of the influenza virus hemagglutinin glycoprotein , 1985, Cell.

[48]  K. Tan,et al.  Atomic structure of a thermostable subdomain of HIV-1 gp41. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[49]  I D Kuntz,et al.  Inhibition of the fusion-inducing conformational change of influenza hemagglutinin by benzoquinones and hydroquinones. , 1993, Biochemistry.

[50]  P. S. Kim,et al.  A spring-loaded mechanism for the conformational change of influenza hemagglutinin , 1993, Cell.

[51]  I. Wilson,et al.  Changes in the conformation of influenza virus hemagglutinin at the pH optimum of virus-mediated membrane fusion. , 1982, Proceedings of the National Academy of Sciences of the United States of America.

[52]  J. Skehel,et al.  The structure of a membrane fusion mutant of the influenza virus haemagglutinin. , 1990, The EMBO journal.

[53]  H. Klenk,et al.  Inhibition of proteolytic activation of influenza virus hemagglutinin by specific peptidyl chloroalkyl ketones , 1989, Virology.

[54]  H. Klenk,et al.  The Molecular Biology of Influenza Virus Pathogenicity , 1988, Advances in Virus Research.