title: Trypsin-resistant protease activation mutants of an influenza virus.
abstract: New classes of mutants of influenza virus A/seal/Mass/1/80 are described in which the haemagglutinins (HA) have lost their protease cleavability by trypsin, but can be activated by elastase, chymotrypsin or thermolysin in different cell types. The same proteases that were required for activation of infectivity of the mutants also activated haemolysis and cell-fusing properties. The protease activation (pa)-mutants were non-pathogenic for chickens, but induced a protective immune response against a highly pathogenic challenge virus. The failure of the mutants to be activated by trypsin, but instead to be activated by the other proteases employed, was related to amino acid exchanges around the HA cleavage site. The cleavability of the chymotrypsin and elastase pa-mutants is most likely determined by replacement of Arg-1 by neutral amino acids such as Ile, Thr, Met or Leu, depending on the substrate specificity of the activating proteases. Cleavage activation of the thermolysin pa-mutants, on the other hand, became possible by insertion of a single Leu residue at position 4 of the HA2 polypeptide, which compensates for the loss of the Gly residue at the N terminus of the fusion peptide due to thermolysin cleavage. The correction of the mutations in revertants confirmed the conclusions drawn from sequence analyses of the pa-mutants.
authors.affiliation: Institut für Virologie, Justus-Liebig-Universität Giessen, Germany.
authors.forename: M
authors.initials: M
authors.lastname: Orlich
authors.forename: D
authors.initials: D
authors.lastname: Linder
authors.forename: R
authors.initials: R
authors.lastname: Rott
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002459:Cell Fusion; D002642:Chick Embryo; D002645:Chickens; D004252:DNA Mutational Analysis; D010450:Endopeptidases / Q000378:metabolism*; D004789:Enzyme Activation; D006389:Hemagglutinins, Viral / Q000235:genetics / Q000378:metabolism*; D006461:Hemolysis; D007110:Immunity, Active; D009980:Influenza A virus / Q000201:enzymology / Q000235:genetics / Q000472:pathogenicity*; D008969:Molecular Sequence Data; D009154:Mutation / Q000502:physiology*; D009976:Orthomyxoviridae Infections / Q000276:immunology; D046048:Seals, Earless; D017421:Sequence Analysis; D013379:Substrate Specificity; D014357:Trypsin / Q000378:metabolism; D010948:Viral Plaque Assay
chemical_list: D006389:Hemagglutinins, Viral; D010450:Endopeptidases; D014357:Trypsin