HEMADSORPTION AND FUSION INHIBITION ACTIVITIES OF HEMAGGLUTININ ANALYZED BY VACCINIA VIRUS MUTANTS
HEMADSORPTION AND FUSION INHIBITION ACTIVITIES OF HEMAGGLUTININ ANALYZED BY VACCINIA VIRUS MUTANTS
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DOI：
10.1016/0042-6822(90)90422-n
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发表时间：
1990-04-01
期刊：
影响因子：
3.7
通讯作者：
SHIDA, H
中科院分区：
文献类型：
--
作者：
SEKI, M;OIE, M;SHIDA, H
痘苗病毒IHD-J株可诱导感染细胞表面的血凝素(HA)，但不能诱导细胞-细胞融合(F-)。我们从IHD-J中分离到21个独立的血吸附阴性(HAD-)突变体病毒，并从其中一个突变体中分离到5个HAD+突变株。在21个突变体中，19个在细胞表面合成或少量合成HA的突变体引起细胞-细胞融合(F+)，而在细胞表面合成HA的5个回复突变体均未诱导细胞-细胞融合。此外，抗HA单抗B2D10诱导了IHD-J感染细胞的广泛多核细胞增殖，并抑制了IHD-J感染细胞提取物抑制IHD-W诱导的多核细胞增殖的能力。在21个HAD-突变体中，B1和A2分别表现为F-表型和F+表型。突变株B1的HA分子胞外区以Lys取代Glu-121，突变体A2的HA分子以Tyr取代Cys-103。我们的结论是，痘苗病毒HA是一种融合抑制蛋白，这两种活性的活性部位分别位于其胞外区，半胱氨酸-103对形成正确的蛋白质三级结构具有重要作用。
Vaccinia virus IHD-J 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 IHD-W. 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 substituiton 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.