Multiple components of the nuclear pore complex interact with the amino-terminus of MX2 to facilitate HIV-1 restriction.
Multiple components of the nuclear pore complex interact with the amino-terminus of MX2 to facilitate HIV-1 restriction.
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DOI：
10.1371/journal.ppat.1007408
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发表时间：
2018-11
期刊：
影响因子：
6.7
通讯作者：
Malim MH
中科院分区：
文献类型：
--
作者：
Dicks MDJ;Betancor G;Jimenez-Guardeño JM;Pessel-Vivares L;Apolonia L;Goujon C;Malim MH
人粘病毒耐药性2（MX2/MXB）是一种干扰素诱导的1型人免疫缺陷病毒（HIV-1）感染的进入后抑制剂。虽然病毒抑制的确切机制仍不清楚，但MX2定位于核膜，并阻断病毒cDNA的核输入。MX2（N-MX2）的氨基末端对于抗病毒功能至关重要，并且在残基11至13处的三重精氨酸基序的突变消除了抗HIV-1活性。在这项研究中，我们试图通过酵母双杂交筛选鉴定功能相关的宿主编码的相互作用伴侣，以研究N-MX2在抗病毒活性中的作用。值得注意的是，7个主要候选相互作用物中有5个是核孔蛋白或核孔蛋白样蛋白，尽管在用突变RRR 11 - 13 A N-MX2片段筛选时没有鉴定出这些候选物。通过免疫共沉淀证实了相互作用，细胞系和原代CD 4 + T细胞中的RNA沉默实验表明，核孔复合物和核输入机制的多种组分可影响MX2抗病毒活性。特别是，含有细胞质丝核孔蛋白NUP 214和转运受体转运蛋白-1（TNPO 1）的苯丙氨酸-甘氨酸（FG）重复序列始终是完整的MX2和干扰素介导的抗病毒功能所必需的。这两种蛋白质被证明与三重精氨酸基序相互作用，共聚焦荧光显微镜显示，它们的同时耗尽导致减少MX2积累在核膜。因此，我们提出了一个模型，其中核输入机制和核孔复合物的多个组件帮助将MX2定位在核膜上，以促进MX2介导的HIV-1限制。大分子进入细胞核的运动在被称为核孔的核膜内的特定位点受到调节。为了有效地感染细胞，人类免疫缺陷病毒1型（HIV-1）必须穿过核膜，使病毒DNA整合到宿主细胞的基因组DNA中。我们和其他人先前已经确定了一种细胞编码的蛋白质，即人类粘病毒抗性2（MX2），它在先天免疫应答启动时表达，并防止HIV-1 DNA在细胞核内积累，从而阻止感染。在这里，我们揭示了核孔复合物的成分，和核进口机械，需要MX2依赖性抑制HIV-1感染。我们发现，MX2，这是本地化的核膜的细胞质面，与核孔复合物的多个蛋白质组分，以及转运受体转运蛋白-1，通过功能上需要的三重精氨酸基序在其氨基末端相互作用。我们推测，这些相互作用促进MX2介导的抑制HIV-1的核输入位于核膜蛋白。
Human myxovirus resistance 2 (MX2/MXB) is an interferon-induced post-entry inhibitor of human immunodeficiency virus type-1 (HIV-1) infection. While the precise mechanism of viral inhibition remains unclear, MX2 is localized to the nuclear envelope, and blocks the nuclear import of viral cDNAs. The amino-terminus of MX2 (N-MX2) is essential for anti-viral function, and mutation of a triple arginine motif at residues 11 to 13 abrogates anti-HIV-1 activity. In this study, we sought to investigate the role of N-MX2 in anti-viral activity by identifying functionally relevant host-encoded interaction partners through yeast-two-hybrid screening. Remarkably, five out of seven primary candidate interactors were nucleoporins or nucleoporin-like proteins, though none of these candidates were identified when screening with a mutant RRR11-13A N-MX2 fragment. Interactions were confirmed by co-immunoprecipitation, and RNA silencing experiments in cell lines and primary CD4+ T cells demonstrated that multiple components of the nuclear pore complex and nuclear import machinery can impact MX2 anti-viral activity. In particular, the phenylalanine-glycine (FG) repeat containing cytoplasmic filament nucleoporin NUP214, and transport receptor transportin-1 (TNPO1) were consistently required for full MX2, and interferon-mediated, anti-viral function. Both proteins were shown to interact with the triple arginine motif, and confocal fluorescence microscopy revealed that their simultaneous depletion resulted in diminished MX2 accumulation at the nuclear envelope. We therefore propose a model whereby multiple components of the nuclear import machinery and nuclear pore complex help position MX2 at the nuclear envelope to promote MX2-mediated restriction of HIV-1. The movement of large molecules into the cell nucleus is regulated at specific sites within the nuclear envelope termed nuclear pores. To infect cells productively, human immunodeficiency virus type-1 (HIV-1) must traverse the nuclear envelope to enable integration of the viral DNA into the genomic DNA of host cells. We, and others, have previously identified a cell-encoded protein, human myxovirus resistance 2 (MX2), which is expressed upon initiation of an innate immune response and prevents accumulation of HIV-1 DNA within the nucleus, thus imposing a block to infection. Here, we reveal that components of the nuclear pore complex, and nuclear import machinery, are required for MX2-dependent inhibition of HIV-1 infection. We show that MX2, which is localized at the cytoplasmic face of the nuclear envelope, interacts with multiple protein components of the nuclear pore complex, as well as transport receptor transportin-1, via a functionally required triple arginine motif at its amino-terminus. We speculate that these interactions facilitate MX2-mediated inhibition of HIV-1 nuclear import by situating the protein at the nuclear envelope.
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影响因子：
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作者：
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通讯作者：
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DOI：
10.1073/pnas.0403167101
发表时间：
2004-06-15
影响因子：
11.1
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通讯作者：
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