Chromosomal integration of LTR-flanked DNA in yeast expressing HIV-1 integrase: down regulation by RAD51.
Chromosomal integration of LTR-flanked DNA in yeast expressing HIV-1 integrase: down regulation by RAD51.
复制标题
DOI：
10.1093/nar/gkl843
复制
发表时间：
2006
影响因子：
14.9
通讯作者：
Parissi V
中科院分区：
文献类型：
--
作者：
Desfarges S;San Filippo J;Fournier M;Calmels C;Caumont-Sarcos A;Litvak S;Sung P;Parissi V
HIV-1整合酶（IN）是催化前病毒DNA整合的关键酶。虽然该酶在体外准确地催化整合步骤，但IN是否足以用于体内整合以及它如何与细胞机制相互作用仍不清楚。我们建立了一个酵母细胞整合系统，其中整合酶作为唯一的HIV-1蛋白表达并靶向染色体。在这种简单的真核模型中，整合酶对于将含有病毒LTR的DNA插入基因组中是必要的和足够的，从而允许独立于其他病毒机制研究分离的整合步骤。此外，酵母系统被用来确定参与整合步骤的细胞机制，并使我们能够显示同源重组系统的作用。我们证明了HIV-1 IN和RAD 51蛋白之间的物理相互作用，并表明HIV-1整合酶活性可以在细胞内和体外被RAD 51蛋白抑制。我们的数据允许鉴定RAD 51作为一种新的体外IN辅因子，能够下调这种逆转录病毒酶的活性，从而作为HIV感染的潜在细胞限制因子。
HIV-1 integrase (IN) is the key enzyme catalyzing the proviral DNA integration step. Although the enzyme catalyzes the integration step accurately in vitro, whether IN is sufficient for in vivo integration and how it interacts with the cellular machinery remains unclear. We set up a yeast cellular integration system where integrase was expressed as the sole HIV-1 protein and targeted the chromosomes. In this simple eukaryotic model, integrase is necessary and sufficient for the insertion of a DNA containing viral LTRs into the genome, thereby allowing the study of the isolated integration step independently of other viral mechanisms. Furthermore, the yeast system was used to identify cellular mechanisms involved in the integration step and allowed us to show the role of homologous recombination systems. We demonstrated physical interactions between HIV-1 IN and RAD51 protein and showed that HIV-1 integrase activity could be inhibited both in the cell and in vitro by RAD51 protein. Our data allowed the identification of RAD51 as a novel in vitro IN cofactor able to down regulate the activity of this retroviral enzyme, thereby acting as a potential cellular restriction factor to HIV infection.
登录
查看更多内容
DOI：
10.1074/jbc.m505367200
发表时间：
2005-08-12
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Li M;Craigie R
通讯作者：
Craigie R
影响因子：
3.8
作者：
Chi, P;Van Komen, S;Sung, P
通讯作者：
Sung, P
影响因子：
4.8
作者：
Cherepanov, P;Maertens, G;Debyser, Z
通讯作者：
Debyser, Z
DOI：
10.1073/pnas.051633498
发表时间：
2001-03-13
影响因子：
11.1
作者：
Ha, HC;Juluri, K;Snyder, SH
通讯作者：
Snyder, SH
影响因子：
56.9
作者：
KALPANA, GV;MARMON, S;GOFF, SP
通讯作者：
GOFF, SP