HIV-1 Vpu Antagonizes CD317/Tetherin by Adaptor Protein-1-Mediated Exclusion from Virus Assembly Sites
HIV-1 Vpu Antagonizes CD317/Tetherin by Adaptor Protein-1-Mediated Exclusion from Virus Assembly Sites
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HIV-1 Vpu 通过接头蛋白 1 介导的病毒组装位点排斥来拮抗 CD317/Tetherin
DOI：
10.1128/jvi.00504-16
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发表时间：
2016
影响因子：
5.4
通讯作者：
Fackler OT
中科院分区：
文献类型：
--
作者：
Pujol FM;Laketa V;Schmidt F;Mukenhirn M;Müller B;Boulant S;Grimm D;Keppler OT;Fackler OT
宿主细胞限制因子CD317/tetherin将病毒粒子捕获在产生细胞的表面，以防止它们的释放。HIV-1辅助蛋白VPU拮抗这一限制。VPU降低了限制因子的细胞表面密度，并将其用于降解；然而，这些活性对于促进颗粒释放是必不可少的。相反，VPU被认为通过阻止内化的CD317/tetherin循环到细胞表面，阻止新合成的CD317/tetherin的顺行运输，和/或取代质膜上病毒组装部位的限制因子来拮抗CD317/tetherin。在分子水平上，拮抗作用依赖于VPU与CD317/Tetherin的物理相互作用。最近的研究结果表明，一个二甘氨酸基序的磷酸化使VPU能够通过两个独立的相互作用基序与接合蛋白1(AP-1)运输复合体结合，并将CD317/Tetherin偶联到内吞机制。在这里，我们使用了一组在单个相互作用基序中具有特定突变的VPU蛋白来定义CD317/tetherin的拮抗需要哪些相互作用。阻碍CD317/Tetherin向质膜的再循环或顺行转运不足以产生拮抗作用。相反，从HIV-1组装位点排除CD317/tetherin取决于VPU基序与AP-1和CD317/tetherin的相互作用，并与颗粒释放限制的拮抗作用相关。一直以来，对AP-1功能或其表达的干扰阻断了这些VPU活动。我们的结果将HIV-1组装位点的置换定义为VPU拮抗CD317/tetherin的活性原理，并支持VPU、AP-1和CD317/tetherin之间的三方复合物在这一过程中的作用。IMPORTANCECD317/tetherin通过将病毒颗粒捕获在分泌细胞表面，从而阻止病毒颗粒在人类细胞中复制，从而对人类免疫缺陷病毒1型(HIV-1)的复制构成内在障碍。病毒蛋白VPU拮抗这种限制，与限制因子和接头蛋白复合体1(AP-1)的分子相互作用被认为是介导这种活性的机制。VPU调节CD317/Tetherin的细胞内运输，并排除质膜上HIV-1组装位点的限制因子，但这些作用对拮抗作用的相对贡献仍不清楚。利用一组VPU突变体，以及对AP-1功能和表达的干扰，我们在这里证明了VPU通过阻止CD317/tetherin以AP-1依赖的方式募集到病毒组装位点来拮抗CD317/tetherin。这些结果加深了我们对CD317/Tetherin拮抗的分子机制的理解，并提示VPU与限制因子和AP-1的复合物是潜在的治疗干预的靶点。
The host cell restriction factor CD317/tetherin traps virions at the surface of producer cells to prevent their release. The HIV-1 accessory protein Vpu antagonizes this restriction. Vpu reduces the cell surface density of the restriction factor and targets it for degradation; however, these activities are dispensable for enhancing particle release. Instead, Vpu has been suggested to antagonize CD317/tetherin by preventing recycling of internalized CD317/tetherin to the cell surface, blocking anterograde transport of newly synthesized CD317/tetherin, and/or displacing the restriction factor from virus assembly sites at the plasma membrane. At the molecular level, antagonism relies on the physical interaction of Vpu with CD317/tetherin. Recent findings suggested that phosphorylation of a diserine motif enables Vpu to bind to adaptor protein 1 (AP-1) trafficking complexes via two independent interaction motifs and to couple CD317/tetherin to the endocytic machinery. Here, we used a panel of Vpu proteins with specific mutations in individual interaction motifs to define which interactions are required for antagonism of CD317/tetherin. Impairing recycling or anterograde transport of CD317/tetherin to the plasma membrane was insufficient for antagonism. In contrast, excluding CD317/tetherin from HIV-1 assembly sites depended on Vpu motifs for interaction with AP-1 and CD317/tetherin and correlated with antagonism of the particle release restriction. Consistently, interference with AP-1 function or its expression blocked these Vpu activities. Our results define displacement from HIV-1 assembly sites as active principle of CD317/tetherin antagonism by Vpu and support a role of tripartite complexes between Vpu, AP-1, and CD317/tetherin in this process.IMPORTANCECD317/tetherin poses an intrinsic barrier to human immunodeficiency virus type 1 (HIV-1) replication in human cells by trapping virus particles at the surface of producer cells and thereby preventing their release. The viral protein Vpu antagonizes this restriction, and molecular interactions with the restriction factor and adaptor protein complex 1 (AP-1) were suggested to mediate this activity. Vpu modulates intracellular trafficking of CD317/tetherin and excludes the restriction factor from HIV-1 assembly sites at the plasma membrane, but the relative contribution of these effects to antagonism remain elusive. Using a panel of Vpu mutants, as well as interference with AP-1 function and expression, we show here that Vpu antagonizes CD317/tetherin by blocking its recruitment to viral assembly sites in an AP-1-dependent manner. These results refine our understanding of the molecular mechanisms of CD317/tetherin antagonism and suggest complexes of Vpu with the restriction factor and AP-1 as targets for potential therapeutic intervention.
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影响因子：
6.7
作者：
Ivanchenko S;Godinez WJ;Lampe M;Kräusslich HG;Eils R;Rohr K;Bräuchle C;Müller B;Lamb DC
通讯作者：
Lamb DC
影响因子：
5.4
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通讯作者：
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影响因子：
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通讯作者：
Kirchhoff F
影响因子：
11.8
作者：
Guo X;Mattera R;Ren X;Chen Y;Retamal C;González A;Bonifacino JS
通讯作者：
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影响因子：
3.7
作者：
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通讯作者：
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