Hemadsorption and fusion inhibition activities of hemagglutinin analyzed by vaccinia virus mutants
| Autor: | Masayasu Oie, Yasuo Ichihashi, Hisatoshi Shida, Makoto Seki |
|---|---|
| Rok vydání: | 1990 |
| Předmět: |
Mutant
Immunoblotting Hemagglutinin (influenza) Fluorescent Antibody Technique Hemagglutinins Viral Vaccinia virus Virus Cell Fusion Virology Extracellular Animals Humans Poxviridae Hemadsorption Orthopoxvirus Vero Cells Cell fusion biology Antibodies Monoclonal biology.organism_classification Molecular biology Precipitin Tests Protein tertiary structure Phenotype Mutation biology.protein HeLa Cells |
| Zdroj: | Virology. 175(2) |
| ISSN: | 0042-6822 |
| Popis: | Vaccinia virus IHDA strain induces hemagglutinin (HA) on the surface membrane of infected cells and does not elicit cell-cell fusion (F − ). We isolated 21 independent hemadsorption-negative (HAD − ) mutant viruses from IHD-J and five HAD + revertants from one of these mutants. Of the 21 mutants, 19 that synthesized either no or little HA at the cell surface caused cell-cell fusion (F + ), whereas none of the five revertants that synthesized HA at the cell surface induced cell-cell fusion. Furthermore, anti-HA monoclonal antibody B2D10 induced extensive polykaryocytosis of IHD-J-infected cells and suppressed the ability of the IHD-J-infected cell extract to inhibit the polykaryocytosis induced by IHDW. The other 2 of the 21 HAD − mutants, B1 and A2, which induced HAs at the cell surface, showed F − and F + phenotype, respectively. The HA molecule of mutant B1 had a single amino acid substitution of Lys for Glu-121 in its extracellular domain, whereas that of mutant A2 had a single substitution mutation of Tyr for Cys-103. We conclude that the vaccinia HA is a fusion inhibition protein, that the active sites for the two activities reside separately in its extracellular domain, and that cysteine-103 is important in forming the proper tertiary structure of the protein to exert both activities. |
| Databáze: | OpenAIRE |
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