Four-tiered π interaction at the dimeric interface of HIV-1 integrase critical for DNA integration and viral infectivity
Four-tiered π interaction at the dimeric interface of HIV-1 integrase critical for DNA integration and viral infectivity
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DOI：
10.1016/j.virol.2008.04.030
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发表时间：
2008-08-01
期刊：
影响因子：
3.7
通讯作者：
Neamati, Nouri
中科院分区：
文献类型：
--
作者：
Al-Mawsawi, Laith Q.;Hombrouck, Anneleen;Neamati, Nouri
HIV-1整合酶（IN）是病毒感染的必需酶。在这里，我们报告了一个广泛的π电子轨道之间的相互作用的四个氨基酸，W132，M178，F181和F185，位于二聚体界面的IN，是关键的链转移活动。评价了9种不同突变IN蛋白在这些位置的催化作用。尽管3 '-加工活性主要是强的，但每种酶的链转移活性完全依赖于二聚体界面处的完整π电子轨道相互作用。在感染性NL4.3 HIV-1病毒克隆的背景下构建了四种代表性IN突变体。而在IN二聚体界面处具有完整的pi电子轨道相互作用的病毒复制与野生型相当，含有废除的pi相互作用的病毒是非感染性的。病毒复制过程中的病毒DNA形式的Q-PCR分析揭示了大多数突变的多效性效应。我们假设π相互作用是功能性IN多聚体复合物组装的关键接触点，并且IN多聚化是功能性预整合复合物所需的。以破坏这种π-π相互作用为目标的小分子抑制剂的合理设计应该导致强大的抗逆转录病毒药物。(C)2008年爱思唯尔公司All rights reserved.
HIV-1 integrase (IN) is an essential enzyme for viral infection. Here, we report an extensive pi electron orbital interaction between four amino acids, W132, M178, F181 and F185, located at the dimeric interface of IN that is critical for the strand transfer activity alone. Catalysis of nine different mutant IN proteins at these positions were evaluated. Whereas the 3'-processing activity is predominantly strong, the strand transfer activity of each enzyme was completely dependent on an intact pi electron orbital interaction at the dimeric interface. Four representative IN mutants were constructed in the context of the infectious NL4.3 HIV-1 viral clone. Whereas viruses with an intact pi electron orbital interaction at the IN dimeric interface replicated comparable to wild type, viruses containing an abolished pi interaction were non-infectious. Q-PCR analysis of viral DNA forms during viral replication revealed pleiotropic effects of most mutations. We hypothesize that the pi interaction is a critical contact point for the assembly of functional IN multimeric complexes, and that IN multimerization is required for a functional pre-integration complex. The rational design of small molecule inhibitors targeting the disruption of this pi-pi interaction should lead to powerful anti-retroviral drugs. (C) 2008 Elsevier Inc. All rights reserved.