HIV integration targeting: a pathway involving Transportin-3 and the nuclear pore protein RanBP2.
HIV integration targeting: a pathway involving Transportin-3 and the nuclear pore protein RanBP2.
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DOI：
10.1371/journal.ppat.1001313
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发表时间：
2011-03
期刊：
影响因子：
6.7
通讯作者：
Bushman FD
中科院分区：
文献类型：
--
作者：
Ocwieja KE;Brady TL;Ronen K;Huegel A;Roth SL;Schaller T;James LC;Towers GJ;Young JA;Chanda SK;König R;Malani N;Berry CC;Bushman FD
全基因组siRNA筛选已经鉴定出对有效HIV感染重要的宿主细胞因子，其中包括核孔蛋白如RanBP 2/Nup 358和核粒转运蛋白转运蛋白-3/TNPO 3。对这些蛋白在HIV复制周期中的作用的分析表明，通过孔的正确运输可能有助于随后的整合步骤。在这里，我们通过证明转运蛋白-3或RanBP 2的缺失改变了早期HIV复制的末端步骤，即染色体整合位点的选择，来展示这些步骤之间的耦合数据。我们发现转运蛋白-3和RanBP 2的缺失改变了HIV的整合靶向。这些敲除降低了HIV在基因密集区和基因相关特征附近的整合频率，这一模式不同于HIV整合酶结合辅因子Psip 1/Ledgf/p75的耗竭。MLV整合不受转运蛋白-3敲低的影响。使用siRNA敲除和整合靶向分析，我们还涉及几个额外的核蛋白在适当的靶位点选择。为了绘制整合靶向的病毒决定簇，我们分析了含有MLV gag的嵌合HIV衍生物，发现gag替代表型模仿了转运蛋白-3和RanBP 2敲低。因此，我们的数据支持一个模型，其中适当的运输和核孔机械介导的GAG依赖性参与贩运的HIV复合物的网站的整合。艾滋病毒每年继续造成全世界约200万人死亡。作为病毒复制周期的一部分，病毒cDNA通过核孔转运到细胞核中，在那里它整合到宿主细胞基因组中。HIV整合是非随机的，可能选择宿主染色体内最能使病毒基因表达并最终产生子代病毒的整合位点。HIV利用宿主因素来指导其整合位点的选择。在这里，我们表明，核贩运和核孔机制的组成部分是艾滋病毒实现其正常模式的整合网站。这一发现表明，病毒通过核孔进入细胞核与下游整合事件相关联，并使病毒能够在宿主基因组中达到其最终位置。我们的研究为HIV复制周期的两个重要步骤提供了新的见解，并为抗逆转录病毒药物提供了可能的新靶点。
Genome-wide siRNA screens have identified host cell factors important for efficient HIV infection, among which are nuclear pore proteins such as RanBP2/Nup358 and the karyopherin Transportin-3/TNPO3. Analysis of the roles of these proteins in the HIV replication cycle suggested that correct trafficking through the pore may facilitate the subsequent integration step. Here we present data for coupling between these steps by demonstrating that depletion of Transportin-3 or RanBP2 altered the terminal step in early HIV replication, the selection of chromosomal sites for integration. We found that depletion of Transportin-3 and RanBP2 altered integration targeting for HIV. These knockdowns reduced HIV integration frequency in gene-dense regions and near gene-associated features, a pattern that differed from that reported for depletion of the HIV integrase binding cofactor Psip1/Ledgf/p75. MLV integration was not affected by the Transportin-3 knockdown. Using siRNA knockdowns and integration targeting analysis, we also implicated several additional nuclear proteins in proper target site selection. To map viral determinants of integration targeting, we analyzed a chimeric HIV derivative containing MLV gag, and found that the gag replacement phenocopied the Transportin-3 and RanBP2 knockdowns. Thus, our data support a model in which Gag-dependent engagement of the proper transport and nuclear pore machinery mediate trafficking of HIV complexes to sites of integration. HIV continues to be responsible for approximately two million deaths worldwide each year. As part of the viral replication cycle, the viral cDNA is transported through the nuclear pore into the nucleus where it integrates into the host cell genome. HIV integrates non-randomly, likely choosing integration sites within the host chromosomes that best enable the viral genes to be expressed and, ultimately, progeny virus to be produced. HIV uses host factors to guide its selection of integration sites. Here we demonstrate that components of the nuclear trafficking and nuclear pore machinery are required for HIV to achieve its normal pattern of integration sites. This finding suggests that passage of the virus through the nuclear pore into the nucleus is coupled to downstream integration events and enables the virus to achieve its final position within the host genome. Our study provides new insights into two important steps of the HIV replication cycle and suggests possible new targets for anti-retroviral drugs.
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影响因子：
4.8
作者：
Cherepanov, P;Maertens, G;Debyser, Z
通讯作者：
Debyser, Z
逆转录病毒DNA整合：靶位点选择的病毒和细胞决定因素。
影响因子：
6.7
作者：
Lewinski, Mary K;Yamashita, Masahiro;Emerman, Michael;Ciuffi, Angela;Marshall, Heather;Crawford, Gregory;Collins, Francis;Shinn, Paul;Leipzig, Jeremy;Hannenhalli, Sridhar;Berry, Charles C;Ecker, Joseph R;Bushman, Frederic D
通讯作者：
Bushman, Frederic D
影响因子：
30.3
作者：
Lee K;Ambrose Z;Martin TD;Oztop I;Mulky A;Julias JG;Vandegraaff N;Baumann JG;Wang R;Yuen W;Takemura T;Shelton K;Taniuchi I;Li Y;Sodroski J;Littman DR;Coffin JM;Hughes SH;Unutmaz D;Engelman A;KewalRamani VN
通讯作者：
KewalRamani VN
影响因子：
4.8
作者：
Emiliani, S;Mousnier, A;Benarous, R
通讯作者：
Benarous, R
影响因子：
10.5
作者：
Brady, Troy;Lee, Young Nam;Bushman, Frederic D.
通讯作者：
Bushman, Frederic D.