Mutations at human immunodeficiency virus type 1 reverse transcriptase tryptophan repeat motif attenuate the inhibitory effect of efavirenz on virus production
Mutations at human immunodeficiency virus type 1 reverse transcriptase tryptophan repeat motif attenuate the inhibitory effect of efavirenz on virus production
复制标题
DOI：
10.1016/j.virol.2008.10.027
复制
发表时间：
2009-01-20
期刊：
影响因子：
3.7
通讯作者：
Wang, Chin-Tien
中科院分区：
文献类型：
--
作者：
Chiang, Chien-Cheng;Wang, Shiu-Mei;Wang, Chin-Tien
HIV-1病毒颗粒加工由蛋白酶（PR）介导，酶促活化由Gag-Pol/Gag-Pol相互作用触发。我们以前报道过，截短突变的逆转录酶（RT）连接子域显着损害病毒颗粒的处理，这表明PR介导的病毒处理的RT子域的重要作用。HIV-1 RT连接亚结构域的一个高度保守的色氨酸（Trp）重复基序参与RT二聚化。本研究的目的是确定Trp重复基序的突变是否对PR介导的病毒加工有任何影响。我们的结果表明，尽管W 401（W 401 A）或W 401和W 402（W 401 A/W 402 A）处的丙氨酸取代对稳态病毒加工没有重大影响，但组合的W 401 A/W 402 A突变部分抵消了依非韦伦（EFV）对病毒产生的屏障。RT不稳定性和差的酶活性的组合反映了由突变引起的RT二聚化缺陷。我们还发现，携带W 401 A或W 401 A/W 402 A突变的人工p66 RT比野生型p66 RT更有效地包装到病毒体中，并且EFV显著降低了p66 RT的病毒掺入，这意味着EFV对RT-Gag相互作用的新作用。我们的研究结果表明，色氨酸重复基序可能在Gag-Pol/Gag-Pol相互作用中发挥作用，有助于随后的PR激活。(c)2008年爱思唯尔公司All rights reserved.
HIV-1 virus particle processing is mediated by protease (PR), with enzymatic activation triggered by Gag-Pol/Gag-Pol interaction. We previously reported that truncation mutations at the reverse transcriptase (RT) connection subdomain markedly impair virus particle processing, suggesting an important role for the RT subdomain in PR-mediated virus processing. A highly conserved tryptophan (Trp) repeat motif of the HIV-1 RT connection subdomain is involved in RT dimerization. Our goal in this study was to determine whether mutations at the Trp repeat motif have any effect on PR-mediated virus processing. Our results indicate that even though alanine substitutions at W401 (W401 A) or at both W401 and W402 (W401A/W402A) have no major effect on steady-state virus processing, the combined W401A/W402A mutations partially negate and the W401 A mutation almost completely negates an efavirenz (EFV)-imposed barrier to virus production. The combination of RT instability and poor enzymatic activity reflects a RT dimerization defect incurred by the mutations. We also found that an artificial p66RT carrying the W401A or W401A/W402A Mutations was packaged into virions more efficiently than wild-type p66RT and that the viral incorporation of p66RT is significantly reduced by EFV, implying a novel effect of EFV on RT-Gag interaction. Our results suggest that the Trp repeat motif may play a role in the Gag-Pol/Gag-Pol interaction that contributes to subsequent PR activation. (c) 2008 Elsevier Inc. All rights reserved.