Department of Pathology, Microbiology and Immunology, Vanderbilt University Medical Center, Nashville, Tennessee, USA.
Department of Pathology, Microbiology and Immunology, Vanderbilt University Medical Center, Nashville, Tennessee, USA
J Virol. 2018 Sep 26;92(20). doi: 10.1128/JVI.00358-18. Print 2018 Oct 15.
Human immunodeficiency virus type 1 (HIV-1) infection depends on efficient intracytoplasmic transport of the incoming viral core to the target cell nucleus. Evidence suggests that this movement is facilitated by the microtubule motor dynein, a large multiprotein complex that interacts with dynactin and cargo-specific adaptor proteins for retrograde movement via microtubules. Dynein adaptor proteins are necessary for activating dynein movement and for linking specific cargoes to dynein. We hypothesized that HIV-1 engages the dynein motor complex via an adaptor for intracellular transport. Here, we show that small interfering RNA depletion of the dynein heavy chain, components of the dynactin complex, and the dynein adaptor BICD2 reduced cell permissiveness to HIV-1 infection. Cell depletion of dynein heavy chain and BICD2 resulted in impaired HIV-1 DNA accumulation in the nucleus and decreased retrograde movement of the virus. Biochemical studies revealed that dynein components and BICD2 associate with capsid-like assemblies of the HIV-1 CA protein in cell extracts and that purified recombinant BICD2 binds to CA assemblies Association of dynein with CA assemblies was reduced upon immunodepletion of BICD2 from cell extracts. We conclude that BICD2 is a capsid-associated dynein adaptor utilized by HIV-1 for transport to the nucleus. During HIV-1 infection, the virus must travel across the cytoplasm to enter the nucleus. The host cell motor protein complex dynein has been implicated in HIV-1 intracellular transport. We show that expression of the dynein heavy chain, components of the dynein-associated dynactin complex, and the dynein adaptor BICD2 in target cells are important for HIV-1 infection and nuclear entry. BICD2 interacts with the HIV-1 capsid , suggesting that it functions as a capsid-specific adaptor for HIV-1 intracellular transport. Our work identifies specific host proteins involved in microtubule-dependent HIV-1 intracellular transport and highlights the BICD2-capsid interaction as a potential target for antiviral therapy.
人类免疫缺陷病毒 1 型（HIV-1）感染依赖于将传入的病毒核心高效地运输到靶细胞的细胞核内。有证据表明，这种运动是由微管动力蛋白 dynein 促进的，dynein 是一种大型多蛋白复合物，与 dynactin 相互作用，并通过微管进行逆行运动。dynein 衔接蛋白对于激活 dynein 运动和将特定货物连接到 dynein 是必需的。我们假设 HIV-1 通过细胞内运输的衔接蛋白与 dynein 运动复合物结合。在这里，我们表明，dynein 重链、dynactin 复合物的成分和 dynein 衔接蛋白 BICD2 的小干扰 RNA 耗竭降低了细胞对 HIV-1 感染的易感性。dynein 重链和 BICD2 的细胞耗竭导致 HIV-1 病毒 DNA 在细胞核内的积累受损，病毒的逆行运动减少。生化研究表明，dynein 成分和 BICD2 与 HIV-1 CA 蛋白的衣壳样组装体在细胞提取物中相互作用，并且纯化的重组 BICD2 与 CA 组装体结合。在从细胞提取物中免疫耗竭 BICD2 后，dynein 与 CA 组装体的结合减少。我们得出结论，BICD2 是 HIV-1 用于运输到细胞核的衣壳相关 dynein 衔接蛋白。在 HIV-1 感染过程中，病毒必须穿过细胞质进入细胞核。宿主细胞的马达蛋白复合物 dynein 已被牵连到 HIV-1 的细胞内运输中。我们表明，靶细胞中 dynein 重链、dynein 相关 dynactin 复合物的成分和 dynein 衔接蛋白 BICD2 的表达对 HIV-1 感染和核进入是重要的。BICD2 与 HIV-1 衣壳相互作用，表明它作为 HIV-1 细胞内运输的衣壳特异性衔接蛋白发挥作用。我们的工作确定了参与微管依赖性 HIV-1 细胞内运输的特定宿主蛋白，并强调了 BICD2-衣壳相互作用作为抗病毒治疗的潜在靶点。
