The AP-1 binding sites located in the pol gene intragenic regulatory region of HIV-1 are important for viral replication.
The AP-1 binding sites located in the pol gene intragenic regulatory region of HIV-1 are important for viral replication.
复制标题
DOI：
10.1371/journal.pone.0019084
复制
发表时间：
2011-04-19
期刊：
影响因子：
3.7
通讯作者：
Van Lint C
中科院分区：
文献类型：
--
作者：
Colin L;Vandenhoudt N;de Walque S;Van Driessche B;Bergamaschi A;Martinelli V;Cherrier T;Vanhulle C;Guiguen A;David A;Burny A;Herbein G;Pancino G;Rohr O;Van Lint C
我们的实验室先前已经在人类免疫缺陷病毒1型（HIV-1）基因组中发现了一个重要的基因内区域，其完整的功能单元由5103片段、dnasei -超敏位点HS7和5105片段组成。这些片段（5103和5105）对单纯疱疹病毒胸苷激酶启动子均表现出PMA诱导的增强活性。在这里，我们通过展示pma诱导的c-Fos、JunB和JunD与位于pol基因末端的片段的体外结合和体内募集，表征了先前鉴定的片段5103的三个AP-1结合位点。功能分析表明，基因内AP-1结合位点完全负责片段5103的pma依赖性增强子活性。此外，用野生型和突变型病毒感染t淋巴Jurkat和前单核细胞U937细胞表明，基因内AP-1位点的突变单独或联合改变了HIV-1的复制。重要的是，三个基因内AP-1位点的突变导致RNA聚合酶II在体内募集到病毒启动子的减少，这有力地支持了这些突变对病毒复制的有害影响至少部分发生在转录水平上。单核细胞源性巨噬细胞的单轮感染证实了基因内AP-1位点对HIV-1感染的重要性。
Our laboratory has previously identified an important intragenic region in the human immunodeficiency virus type 1 (HIV-1) genome, whose complete functional unit is composed of the 5103 fragment, the DNaseI-hypersensitive site HS7 and the 5105 fragment. These fragments (5103 and 5105) both exhibit a phorbol 12-myristate 13-acetate (PMA)-inducible enhancer activity on the herpes simplex virus thymidine kinase promoter. Here, we characterized the three previously identified AP-1 binding sites of fragment 5103 by showing the PMA-inducible in vitro binding and in vivo recruitment of c-Fos, JunB and JunD to this fragment located at the end of the pol gene. Functional analyses demonstrated that the intragenic AP-1 binding sites are fully responsible for the PMA-dependent enhancer activity of fragment 5103. Moreover, infection of T-lymphoid Jurkat and promonocytic U937 cells with wild-type and mutant viruses demonstrated that mutations of the intragenic AP-1 sites individually or in combination altered HIV-1 replication. Importantly, mutations of the three intragenic AP-1 sites led to a decreased in vivo recruitment of RNA polymerase II to the viral promoter, strongly supporting that the deleterious effect of these mutations on viral replication occurs, at least partly, at the transcriptional level. Single-round infections of monocyte-derived macrophages confirmed the importance of intragenic AP-1 sites for HIV-1 infectivity.
登录
查看更多内容
影响因子：
4
作者：
Hess, J;Angel, P;Schorpp-Kistner, M
通讯作者：
Schorpp-Kistner, M
影响因子：
4.8
作者：
Basbous, J;Arpin, C;Mesnard, JM
通讯作者：
Mesnard, JM
影响因子：
14.9
作者：
Goffin, W;Demonté, D;Van Lint, C
通讯作者：
Van Lint, C
影响因子：
5.4
作者：
HARRICH, D;HSU, C;GAYNOR, RB
通讯作者：
GAYNOR, RB
影响因子：
5.4
作者：
Inoshima, Y;Kohmoto, M;Mikami, T
通讯作者：
Mikami, T