Biochemical and genetic analyses of integrase-interacting proteins lens epithelium-derived growth factor growth factor related protein 2 function and (LEDGF)/p75 and hepatoma-derived (HRP2) in preintegration complex HTV-1 replication
Biochemical and genetic analyses of integrase-interacting proteins lens epithelium-derived growth factor growth factor related protein 2 function and (LEDGF)/p75 and hepatoma-derived (HRP2) in preintegration complex HTV-1 replication
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DOI：
10.1016/j.virol.2005.11.022
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发表时间：
2006-03-15
期刊：
影响因子：
3.7
通讯作者：
Engelman, A
中科院分区：
文献类型：
--
作者：
Vandegraaff, N;Devroe, E;Engelman, A
人类免疫缺陷病毒1型（HIV-1）整合酶（IN）在高分子量整合前复合物（PIC）的背景下在细胞中发挥作用。透镜上皮衍生生长因子（LEDGF）转录辅激活因子/p75和肝癌衍生生长因子相关蛋白2（HRP 2）与HIV-1 IN紧密结合并刺激其体外整合活性。在这里，我们表明，每个重组宿主细胞因子有效地重建的HIV-1 PTCs的体外活性破坏的功能整合的预处理与高浓度的盐。突变分析表明，LEDGF/p75的IN结合和NA结合活性都有助于功能性PIC重建。我们还研究了这些蛋白质在HIV-1感染中的作用，通过使用短干扰RNA。HIV-1感染在LEDGF/p75、HRP 2或这两种蛋白缺失的HeLa-P4细胞中基本上不受影响。我们的结论是，敲除LEDGF/p75和/或HRP 2的细胞将是有用的遗传工具，以解决这些宿主细胞因子在HIV-1复制中的作用。(C)2005年爱思唯尔公司All rights reserved.
Human immunodeficiency virus type 1 (HIV-1) integrase (IN) functions in cells within the context of high molecular weight preintegration complexes (PICs). Lens epithelium-derived growth factor (LEDGF) transcriptional coactivator/p75 and hepatoma-derived growth factor related protein 2 (HRP2) tightly bind to HIV-1 IN and stimulate its integration activity in vitro. Here, we show that each recombinant host cell factor efficiently reconstitutes the in vitro activity of HIV-1 PTCs disrupted for functional integration by pre-treatment with high concentrations of salt. Mutational analysis reveals that both the IN-binding and I)NA-binding activities of LEDGF/p75 contribute to functional PIC reconstitution. We also investigate a role(s) for these proteins in HIV-1 infection by using short-interfering RNA. HIV-1 infection was essentially unaffected in HeLa-P4 cells depleted for LEDGF/p75, HRP2, or both proteins. We conclude that cells knocked-out for LEDGF/p75 and/or HRP2 will be useful genetic tools to address the roles of these host cell factors in HIV-1 replication. (C) 2005 Elsevier Inc. All rights reserved.