Docking of HIV-1 Vpr to the nuclear envelope is mediated by the interaction with the nucleoporin hCG1
Docking of HIV-1 Vpr to the nuclear envelope is mediated by the interaction with the nucleoporin hCG1
复制标题
DOI：
10.1074/jbc.m207439200
复制
发表时间：
2002-11-22
影响因子：
4.8
通讯作者：
Benichou, S
中科院分区：
文献类型：
--
作者：
Le Rouzic, E;Mousnier, A;Benichou, S
HIV-1基因组包含几个编码辅助蛋白的基因，包括小Vpr蛋白。Vpr影响核膜的完整性，并参与含有病毒DNA的预整合复合体的核易位。在这里，我们通过在表达Vpr-绿色荧光蛋白融合的活细胞上进行的光漂白实验表明，蛋白质在细胞核和细胞质之间穿梭，但很大一部分集中在核膜上，支持Vpr与核孔复合物组分相互作用的假设。在酵母双杂交系统中发现了HIV-1 Vpr与人核孔蛋白CG1 （hCG1）的相互作用，并在体外和转染细胞中得到证实。这种相互作用不涉及hCG1的FG重复结构域，而是涉及蛋白质的n端区域。通过对洋地黄苷渗透细胞的核导入实验，我们证实hCG1参与Vpr在核膜的对接。Vpr与核孔复合物组分的这种关联可能有助于核膜的破坏和病毒DNA的核输入。
The HIV-1 genome contains several genes coding for auxiliary proteins, including the small Vpr protein. Vpr affects the integrity of the nuclear envelope and participates in the nuclear translocation of the preintegration complex containing the viral DNA. Here, we show by photobleaching experiments performed on living cells expressing a Vpr-green fluorescent protein fusion that the protein shuttles between the nucleus and the cytoplasm, but a significant fraction is concentrated at the nuclear envelope, supporting the hypothesis that Vpr interacts with components of the nuclear pore complex. An interaction between HIV-1 Vpr and the human nucleoporin CG1 (hCG1) was revealed in the yeast two-hybrid system, and then confirmed both in vitro and in transfected cells. This interaction does not involve the FG repeat domain of hCG1 but rather the N-terminal region of the protein. Using a nuclear import assay based on digitonin-permeabilized cells, we demonstrate that hCG1 participates in the docking of Vpr at the nuclear envelope. This association of Vpr with a component of the nuclear pore complex may contribute to the disruption of the nuclear envelope and to the nuclear import of the viral DNA.