Nup153 Unlocks the Nuclear Pore Complex for HIV-1 Nuclear Translocation in Nondividing Cells
Nup153 Unlocks the Nuclear Pore Complex for HIV-1 Nuclear Translocation in Nondividing Cells
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DOI：
10.1128/jvi.00648-18
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发表时间：
2018-10-01
影响因子：
5.4
通讯作者：
Diaz-Griffero, Felipe
中科院分区：
文献类型：
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作者：
Buffone, Cindy;Martinez-Lopez, Alicia;Diaz-Griffero, Felipe
人类免疫缺陷病毒1型（HIV-1）显示出感染非分裂细胞的独特能力。HIV-1的衣壳是病毒核输入的病毒决定因子。为了了解HIV-1感染非分裂细胞的能力所涉及的细胞因素，我们试图找到允许病毒感染分裂细胞而不是非分裂细胞的衣壳突变。由于衣壳与核孔蛋白153（Nup 153）的相互作用对于HIV-1的核输入是重要的，因此我们解决了与含有苯丙氨酸-甘氨酸重复（FG重复）的Nup 153衍生肽复合的六聚体HIV-1衣壳的新晶体结构，我们使用Nup 153衍生肽来指导衣壳结合界面的基于结构的诱变。测试了这些衣壳残基突变的HIV-1病毒感染分裂和非分裂细胞的能力。衣壳N57置换的HIV-1病毒感染分裂细胞，但不感染非分裂细胞。有趣的是，具有N57突变的HIV-1病毒进行逆转录，但不进行核转位。突变衣壳也失去了与Nup 153和CPSF 6相互作用的能力。使用小分子PF 74和BI-2阻止了含FG的核孔蛋白（Nups）（如Nup 153）与HIV-1核心的相互作用。对N57突变的HIV-1病毒整合位点的分析显示，以类似于HIV-1在CPSF 6敲除细胞中或HIV-1-N74 D中的方式整合到转录活性基因中减少。N57突变体HIV-1的整合模式可以通过失去衣壳与CPSF 6的相互作用来解释，而衣壳与Nup 153的相互作用是HIV-1感染非分裂细胞所必需的。此外，所观察到的病毒整合概况表明，整合位点的选择是一个多参数的过程，取决于核因子和状态的细胞chromatin.IMPORTANCE区分慢病毒，如HIV-1，从所有其他逆转录病毒的关键优势之一是它能够感染nondividing细胞。HIV-1衣壳与Nup 153和CPSF 6的相互作用对于核进入和整合是重要的;然而，这些蛋白质中的每一个对核输入和整合的贡献尚不清楚。利用遗传学，我们证明了这些蛋白质有助于不同的过程：Nup 153对于非分裂细胞中的HIV-1核输入至关重要，CPSF 6对于HIV-1整合很重要。此外，已知核因子如CPSF 6和染色质状态对于整合位点选择是重要的;然而，影响整合位点选择的优先决定子尚不清楚。这项工作表明，整合位点的选择是一个多参数的过程，取决于核因子和细胞染色质的状态。
Human immunodeficiency virus type 1 (HIV-1) displays the unique ability to infect nondividing cells. The capsid of HIV-1 is the viral determinant for viral nuclear import. To understand the cellular factors involved in the ability of HIV-1 to infect nondividing cells, we sought to find capsid mutations that allow the virus to infect dividing but not nondividing cells. Because the interaction of capsid with the nucleoporin protein 153 (Nup153) is important for nuclear import of HIV-1, we solved new crystal structures of hexameric HIV-1 capsid in complex with a Nup153-derived peptide containing a phenylalanine-glycine repeat (FG repeat), which we used to guide structure-based mutagenesis of the capsid-binding interface. HIV-1 viruses with mutations in these capsid residues were tested for their ability to infect dividing and nondividing cells. HIV-1 viruses with capsid N57 substitutions infected dividing but not nondividing cells. Interestingly, HIV-1 viruses with N57 mutations underwent reverse transcription but not nuclear translocation. The mutant capsids also lost the ability to interact with Nup153 and CPSF6. The use of small molecules PF74 and BI-2 prevented the interaction of FG-containing nucleoporins (Nups), such as Nup153, with the HIV-1 core. Analysis of integration sites in HIV-1 viruses with N57 mutations revealed diminished integration into transcriptionally active genes in a manner resembling that of HIV-1 in CPSF6 knockout cells or that of HIV-1-N74D. The integration pattern of the N57 mutant HIV-1 can be explained by loss of capsid interaction with CPSF6, whereas capsid interaction with Nup153 is required for HIV-1 to infect nondividing cells. Additionally, the observed viral integration profiles suggested that integration site selection is a multiparameter process that depends upon nuclear factors and the state of the cellular chromatin.IMPORTANCE One of the key advantages that distinguish lentiviruses, such as HIV-1, from all other retroviruses is its ability to infect nondividing cells. Interaction of the HIV-1 capsid with Nup153 and CPSF6 is important for nuclear entry and integration; however, the contribution of each of these proteins to nuclear import and integration is not clear. Using genetics, we demonstrated that these proteins contribute to different processes: Nup153 is essential for the HIV-1 nuclear import in nondividing cells, and CPSF6 is important for HIV-1 integration. In addition, nuclear factors such as CPSF6 and the state of the chromatin are known to be important for integration site selection; nevertheless, the preferential determinant influencing integration site selection is not known. This work demonstrates that integration site selection is a multiparameter process that depends upon nuclear factors and the state of the cellular chromatin.