Characterization of human immunodeficiency virus type 1 Pr160gag-pol mutants with truncations downstream of the protease domain
Characterization of human immunodeficiency virus type 1 Pr160gag-pol mutants with truncations downstream of the protease domain
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DOI：
10.1016/j.virol.2004.08.010
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发表时间：
2004-11-10
期刊：
影响因子：
3.7
通讯作者：
Wang, CT
中科院分区：
文献类型：
--
作者：
Liao, WH;Wang, CT
我们通过逐步删除病毒蛋白酶(PR)结构域下游的pol序列，构建了一系列HIV-1 Gag-Pol型突变体。分析了截断突变对病毒颗粒产生和GAG颗粒加工的影响。分析表明，去除整合酶(IN)结构域对颗粒处理的效率没有重大影响，但显著减少了病毒颗粒的萌发。然而，IN和RNaseH结构域的删除使病毒颗粒的产生恢复到野生型水平。P51RT结构域被截断后，病毒颗粒的蛋白降解过程明显受阻。所有截短的Gag-Poll蛋白都能被整合到病毒颗粒中，并表现出与野生型(Wt)相似的免疫荧光染色。我们的数据与以下观点一致，即指导Gag-Poll转运和颗粒掺入的信号是由其N端的Gag结构域决定的。保留完整p51RT的截短Gag-Poll能够补充PR缺陷突变体产生感染性假型病毒粒子，病毒滴度为wt的20-70%，而缺失完整p51RT的Gag-Poll产生的假型病毒粒子不具感染性或感染性较差。这表明，Gag-Poll需要一个完整的p51RT结构域来调节成熟的反式感染病毒颗粒的产生。(C)2004 Elsevier Inc.保留所有权利。
We have constructed a series of HIV-1 Gag-pol mutants by progressive deletion of the pol sequence downstream of the viral protease (PR) domain. Effects of the truncation mutations on virus particle production and Gag particle processing were analyzed. Analysis indicated that removal of the integrase (IN) domain had no major effect on the efficiency of particle processing, but resulted in a marked reduction in virus particle budding. Deletion of both the IN and RNase H domains, however, restored the production of virus particles to wild-type level. The proteolytic processing of virus particle was significantly impaired when the p51RT domain was truncated. All of the truncated Gag-pol proteins could be incorporated into virus particles and demonstrated an immunofluorescence staining pattern similar to that of the wild type (wt). Our data are consistent with the proposal that signals for directing the Gag-pol transport and particle incorporation are determined by its N-terminal Gag domain. Truncated Gag-pol retaining an intact p51RT was able to complement a PR-defective mutant to produce infectious pseudotyped virions, with a virus titer 20-70% of that of wt. Pseudotyped virions produced by the Gag-pol lacking an intact p51RT were noninfectious or poorly infectious. This suggests that an intact p51RT domain is required for the Gag-pol to mediate production of mature infectious virus particles in trans. (C) 2004 Elsevier Inc. All rights reserved.