Conserved amino acids of the human immunodeficiency virus type 2 Vpx nuclear localization signal are critical for nuclear targeting of the viral preintegration complex in non-dividing cells
Conserved amino acids of the human immunodeficiency virus type 2 Vpx nuclear localization signal are critical for nuclear targeting of the viral preintegration complex in non-dividing cells
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DOI：
10.1016/j.virol.2005.10.036
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发表时间：
2006-03-01
期刊：
影响因子：
3.7
通讯作者：
Ratner, L
中科院分区：
文献类型：
--
作者：
Belshan, M;Mahnke, LA;Ratner, L
HIV-2病毒辅助蛋白Vpx与HIV-1的Vpr蛋白相关，但不同。Vpx被包装到病毒体中，并且作为病毒整合前复合物（PIC）的组分，是在非分裂细胞中有效病毒复制所必需的。我们以前曾报道过，Vpx的最小可转移区域，含有亲核性是氨基酸65至72。对不同HIV-2/SIV毒株Vpx序列的分析表明，该区域含有高度保守的氨基酸，包括两个碱性残基（K68，R70）和三个酪氨酸（Y 66，Y 69，Y 71）。在这里，我们表明，突变的基本或酪氨酸残基废除PIC核进口逮捕细胞通过PCR检测病毒整合评估。无细胞病毒的蛋白质印迹检测表明，所有的突变蛋白被纳入病毒粒子，这表明，在被捕的细胞中的复制的缺乏是由于在靶细胞中的Vpx的损失。总之，这些研究绘制了Vpx核定位信号的关键残基，这些残基是有效感染非分裂细胞所需的。(c)2005年爱思唯尔公司All rights reserved.
The HIV-2 viral accessory protein Vpx is related to, but distinct from the Vpr protein of HIV-1. Vpx is packaged into virions and as a component of the viral preintegration complex (PIC) is required for efficient virus replication in non-dividing cells. We have previously reported that the minimal transferable region of Vpx that contained karyophilic properties was aa 65 to 72. Analysis of Vpx sequences from various HIV-2/SIV strains reveals that this region contains highly conserved amino acids, including two basic residues (K68, R70) and three tyrosines (Y66, Y69, Y71). Here, we demonstrate that mutation of the basic or tyrosine residues abolishes PIC nuclear import in arrested cells as assessed by PCR detection of viral integration. Examination of cell-free virus by Western blot indicated that all mutant proteins were incorporated into virions, suggesting that the lack of replication in arrested cells was not due to a loss of Vpx in target cells. Together, these studies map critical residues of the Vpx nuclear localization signal that are required for efficient infection of non-dividing cells. (c) 2005 Elsevier Inc. All rights reserved.