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猫免疫缺陷病毒整合酶突变体的特征分析

Characterization of an integrase mutant of feline immunodeficiency virus.

作者信息

Tomonaga K, Itagaki S I, Kashiwase H, Kawaguchi Y, Inoshima Y, Ikeda Y, Mikami T

机构信息

Department of Veterinary Microbiology, University of Tokyo, Japan.

出版信息

Arch Virol. 1998;143(1):1-14. doi: 10.1007/s007050050263.

Abstract

The role of the integrase region of feline immunodeficiency virus (FIV) in viral replication was examined using an integrase mutant clone of FIV which carries a frameshift mutation in the region. Upon transfection, although the integrase mutant was able to release virus-like particles into the supernatant from the transfected cells, the virions produced by the mutant contained unprocessed gag precursor protein and undetectable levels of reverse transcriptase activity. Furthermore, the mutant virions were unable to direct the synthesis of viral DNA after infection in target cells. To understand this phenotype of the integrase mutant in more detail, we constructed a gag-pol expression plasmid from an FIV molecular clone and assayed roles of the integrase region on virus particle formation following transfection. When an inframe deletion was introduced into the protease region of the expression plasmid, the mutant was able to efficiently release gag- and gag-pol precursor proteins into the supernatant from the transfected cells. An expression plasmid with mutations in both the protease and integrase regions, however, failed to release the gag-pol precursor protein from the cells. These results suggested an essential role for the integrase region for efficient incorporation of the gag-pol precursor into the virions.

摘要

利用猫免疫缺陷病毒(FIV)整合酶区域的一个突变克隆来研究该区域在病毒复制中的作用,此突变克隆在该区域携带一个移码突变。转染后,尽管整合酶突变体能够从转染细胞中释放出病毒样颗粒到上清液中,但该突变体产生的病毒粒子含有未加工的gag前体蛋白且逆转录酶活性水平检测不到。此外,突变病毒粒子在感染靶细胞后无法指导病毒DNA的合成。为了更详细地了解整合酶突变体的这种表型,我们从FIV分子克隆构建了一个gag-pol表达质粒,并检测了整合酶区域在转染后对病毒粒子形成的作用。当在表达质粒的蛋白酶区域引入一个框内缺失时,该突变体能够有效地从转染细胞中释放gag和gag-pol前体蛋白到上清液中。然而,一个在蛋白酶和整合酶区域都有突变的表达质粒未能从细胞中释放gag-pol前体蛋白。这些结果表明整合酶区域对于gag-pol前体有效掺入病毒粒子起着至关重要的作用。

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