The Requirement for Cellular Transportin 3 (TNPO3 or TRN-SR2) during Infection Maps to Human Immunodeficiency Virus Type 1 Capsid and Not Integrase
The Requirement for Cellular Transportin 3 (TNPO3 or TRN-SR2) during Infection Maps to Human Immunodeficiency Virus Type 1 Capsid and Not Integrase
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DOI：
10.1128/jvi.01899-09
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发表时间：
2010-01-01
影响因子：
5.4
通讯作者：
Engelman, Alan
中科院分区：
文献类型：
--
作者：
Krishnan, Lavanya;Matreyek, Kenneth A.;Engelman, Alan
最近的全基因组筛选强调了转运蛋白3在人类免疫缺陷病毒1型（HIV-1）感染和整合前复合体（PIC）核输入中的重要作用。此外，HIV-1整合酶与重组转运蛋白3蛋白相互作用的条件下，莫洛尼鼠白血病病毒（MLV）整合酶未能做到这一点，这表明整合酶转运蛋白3的相互作用可能强调主动逆转录病毒PIC核输入。在这里，我们相关的感染性缺陷，转运蛋白3敲低细胞与体外蛋白结合亲和力的一组扩大的逆转录病毒，包括猿猴免疫缺陷病毒（SIV），牛免疫缺陷病毒（BIV），马传染性贫血病毒（EIAV），猫免疫缺陷病毒（FIV），劳斯肉瘤病毒（RSV），关键解决的作用整合酶-转运蛋白3相互作用的病毒感染。与MLV和RSV相比，慢病毒（FIV除外）显示需要转运蛋白3，产生基于感染的依赖性排序SIV > HIV-1 > BIV和EIAV > MLV、RSV和FIV。相比之下，体外下拉和表面等离子体共振测定确定了明显不同的整合酶-转运蛋白3结合等级：FIV、HIV-1和BIV > SIV和MLV > EIAV。因此，我们的研究结果不能支持整合酶结合在逆转录病毒感染过程中决定转运蛋白3依赖性的关键作用。除了整合酶之外，衣壳也被强调为逆转录病毒核输入决定簇。因此，MLV/HIV-1嵌合体病毒精确定位了致敏的遗传决定因素，从而将转运蛋白3敲低至HIV-1衣壳蛋白。因此，我们得出结论，衣壳，而不是整合酶，是主导的病毒因素，决定运输3依赖性在HIV-1感染。
Recent genome-wide screens have highlighted an important role for transportin 3 in human immunodeficiency virus type 1 (HIV-1) infection and preintegration complex (PIC) nuclear import. Moreover, HIV-1 integrase interacted with recombinant transportin 3 protein under conditions whereby Moloney murine leukemia virus (MLV) integrase failed to do so, suggesting that integrase-transportin 3 interactions might underscore active retroviral PIC nuclear import. Here we correlate infectivity defects in transportin 3 knockdown cells with in vitro protein binding affinities for an expanded set of retroviruses that include simian immunodeficiency virus (SIV), bovine immunodeficiency virus (BIV), equine infectious anemia virus (EIAV), feline immunodeficiency virus (FIV), and Rous sarcoma virus (RSV) to critically address the role of integrase-transportin 3 interactions in viral infection. Lentiviruses, with the exception of FIV, display a requirement for transportin 3 in comparison to MLV and RSV, yielding an infection-based dependency ranking of SIV > HIV-1 > BIV and EIAV > MLV, RSV, and FIV. In vitro pulldown and surface plasmon resonance assays, in contrast, define a notably different integrase-transportin 3 binding hierarchy: FIV, HIV-1, and BIV > SIV and MLV > EIAV. Our results therefore fail to support a critical role for integrase binding in dictating transportin 3 dependency during retrovirus infection. In addition to integrase, capsid has been highlighted as a retroviral nuclear import determinant. Accordingly, MLV/HIV-1 chimera viruses pinpoint the genetic determinant of sensitization to transportin 3 knockdown to the HIV-1 capsid protein. We therefore conclude that capsid, not integrase, is the dominant viral factor that dictates transportin 3 dependency during HIV-1 infection.