Interaction of human immunodeficiency virus type 1 integrase with cellular nuclear import receptor importin 7 and its impact on viral replication
Interaction of human immunodeficiency virus type 1 integrase with cellular nuclear import receptor importin 7 and its impact on viral replication
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DOI：
10.1074/jbc.m610546200
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发表时间：
2007-05-04
影响因子：
4.8
通讯作者：
Yao, Xiaojian
中科院分区：
文献类型：
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作者：
Ao, Zhujun;Huang, Guanyou;Yao, Xiaojian
与所有其他病毒一样，人类免疫缺陷病毒1型(HIV-1)的成功复制在很大程度上依赖于细胞因素。在这项研究中，我们用免疫共沉淀法研究了HIV-1整合酶(IN)与几种宿主核输入因子的相互作用。我们的结果表明，IN在体内与宿主Importin 7(Imp7)特异地相互作用，但不与Importin 8(Imp8)或Importin Alpha(Rch1)相互作用。相反，另一种能够与Rch1结合的HIV-1亲核蛋白MAp17不能与Imp7相互作用，这表明IN和Map17在HIV-1复制过程中可能与不同的细胞途径相互作用。遗传分析表明，IN的C末端结构域是负责IN与Imp7相互作用的区域，IN突变体(K240A、K244A/R263A、K264A)破坏了蛋白质与Imp7的结合能力，表明IN C末端结构域中的(235)WKGPAKLWKG和(262)RRKAK是有效的IN/Imp7相互作用所必需的。利用水疱性口炎病毒G糖蛋白假型HIV单循环复制系统，我们发现IN/Imp7相互作用缺陷突变体不能介导病毒复制，并且在病毒反转录和核输入步骤中都表现出损伤。此外，瞬时敲除HIV-1产生细胞和靶细胞中的Imp7，可使HIV感染受到2.5-3.5倍的抑制。综上所述，我们的结果表明，HIV-1IN与Imp7特异地相互作用，这种病毒/细胞蛋白相互作用有助于有效地感染HIV-1。
Similar to all other viruses, human immunodeficiency virus type 1 ( HIV-1) depends heavily on cellular factors for its successful replication. In this study we have investigated the interaction of HIV-1 integrase (IN) with several host nuclear import factors using co-immunoprecipitation assays. Our results indicate that IN interacts specifically with host importin 7 (Imp7) in vivo, but does not interact with importin 8 (Imp8) or importin alpha(Rch1). In contrast, another HIV-1 karyophilic protein MAp17, which is capable of binding Rch1, fails to interact with Imp7, suggesting that IN and Map17 may interact with different cellular pathways during HIV-1 replication. Genetic analysis revealed that the C-terminal domain of IN is the region responsible for interaction between IN with Imp7, and an IN mutant (K240A, K244A/ R263A, K264A) disrupted the Imp7 binding ability of the protein, indicating that both regions ((235)WKGPAKLLWKG and (262)RRKAK) within the C-terminal domain of IN are required for efficient IN/Imp7 interaction. Using a vesicular stomatitis virus G glycoprotein pseudotyped HIV single-cycle replication system, we showed that the IN/Imp7 interaction-deficient mutant was unable to mediate viral replication and displayed impairment at both viral reverse transcription and nuclear import steps. Moreover, transient knockdown of Imp7 in both HIV-1 producing and target cells resulted in a 2.5 - 3.5-fold inhibition of HIV infection. Altogether, our results indicate that HIV-1 IN specifically interacts with Imp7, and this viral/cellular protein interaction contributes to efficient HIV-1 infection.