Human immunodeficiency virus type 1 Vpr interacts with HHR23A, a cellular protein implicated in nucleotide excision DNA repair.
Human immunodeficiency virus type 1 Vpr interacts with HHR23A, a cellular protein implicated in nucleotide excision DNA repair.
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人类免疫缺陷病毒 1 型 Vpr 与 HHR23A 相互作用，HHR23A 是一种参与核苷酸切除 DNA 修复的细胞蛋白。
DOI：
10.1128/jvi.71.12.9732-9742.1997
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发表时间：
1997
影响因子：
5.4
通讯作者：
Chen,IS
中科院分区：
文献类型：
--
作者：
Withers-Ward,ES;Jowett,JB;Stewart,SA;Xie,YM;Garfinkel,A;Shibagaki,Y;Chow,SA;Shah,N;Hanaoka,F;Sawitz,DG;Armstrong,RW;Souza,LM;Chen,IS
人类免疫缺陷病毒1型（HIV-1）vpr基因是灵长类慢病毒HIV-1、HIV-2和猿猴免疫缺陷病毒中进化上保守的基因。vpr基因产物的独特功能之一是将细胞阻滞在细胞周期的G2期。在这里，我们表明，Vpr物理相互作用与HHR 23 A，一个成员的进化保守的基因家族参与核苷酸切除修复。Vpr与HHR 23 A的相互作用最初通过酵母双杂交筛选鉴定，并通过细菌表达的重组和瞬时表达或化学合成的蛋白质产物之间的直接结合的证明来证实。通过间接免疫荧光和共聚焦显微镜观察HHR 23 A和Vpr表明这两种蛋白质共定位于核膜处或核膜附近。我们还将HHR 23 A中的Vpr结合结构域映射到先前显示与泛素化途径成员具有同源性的蛋白质的C-末端45个氨基酸区域。HHR 23 A和截短的衍生物，其中包括Vpr结合结构域的过表达的结果在部分缓解的G2期阻滞诱导的Vpr，表明Vpr和HHR 23 A之间的相互作用是关键的细胞周期阻滞诱导的Vpr。这些结果进一步支持了Vpr干扰DNA修复过程中所涉及的一种或多种蛋白质的正常功能的假设，因此，干扰了允许细胞从细胞周期的G2期过渡到M期的信号传递。
The human immunodeficiency virus type 1 (HIV-1) vpr gene is an evolutionarily conserved gene among the primate lentiviruses HIV-1, HIV-2, and simian immunodeficiency viruses. One of the unique functions attributed to the vpr gene product is the arrest of cells in the G2 phase of the cell cycle. Here we demonstrate that Vpr interacts physically with HHR23A, one member of an evolutionarily conserved gene family involved in nucleotide excision repair. Interaction of Vpr with HHR23A was initially identified through a yeast two-hybrid screen and was confirmed by the demonstration of direct binding between bacterially expressed recombinant and transiently expressed or chemically synthesized protein products. Visualization of HHR23A and Vpr by indirect immunofluorescence and confocal microscopy indicates that the two proteins colocalize at or about the nuclear membrane. We also map the Vpr-binding domain in HHR23A to a C-terminal 45-amino-acid region of the protein previously shown to have homology to members of the ubiquitination pathway. Overexpression of HHR23A and a truncated derivative which includes the Vpr-binding domain results in a partial alleviation of the G2 arrest induced by Vpr, suggesting that the interaction between Vpr and HHR23A is critical for cell cycle arrest induced by Vpr. These results provide further support for the hypothesis that Vpr interferes with the normal function of a protein or proteins involved in the DNA repair process and, thus, in the transmission of signals that allow cells to transit from the G2 to the M phase of the cell cycle.