Yu Wendong, Ramakrishnan Rajesh, Wang Yan, Chiang Karen, Sung Tzu-Ling, Rice Andrew P
Department of Molecular Virology and Microbiology, Baylor College of Medicine, Houston, Texas, United States of America.
PLoS One. 2008 Sep 5;3(9):e3146. doi: 10.1371/journal.pone.0003146.
HIV-1 is dependent upon cellular co-factors to mediate its replication cycle in CD4(+) T cells and macrophages, the two major cell types infected by the virus in vivo. One critical co-factor is Cyclin T1, a subunit of a general RNA polymerase II elongation factor known as P-TEFb. Cyclin T1 is targeted directly by the viral Tat protein to activate proviral transcription. Cyclin T1 is up-regulated when resting CD4(+) T cells are activated and during macrophage differentiation or activation, conditions that are also necessary for high levels of HIV-1 replication. Because Cyclin T1 is a subunit of a transcription factor, the up-regulation of Cyclin T1 in these cells results in the induction of cellular genes, some of which might be HIV-1 co-factors. Using shRNA depletions of Cyclin T1 and transcriptional profiling, we identified 54 cellular mRNAs that appear to be Cyclin T1-dependent for their induction in activated CD4(+) T Jurkat T cells and during differentiation and activation of MM6 cells, a human monocytic cell line. The promoters for these Cyclin T1-dependent genes (CTDGs) are over-represented in two transcription factor binding sites, SREBP1 and ARP1. Notably, 10 of these CTDGs have been reported to be involved in HIV-1 replication, a significant over-representation of such genes when compared to randomly generated lists of 54 genes (p value<0.00021). The results of siRNA depletion and dominant-negative protein experiments with two CTDGs identified here, CDK11 and Casein kinase 1 gamma 1, suggest that these genes are involved either directly or indirectly in HIV-1 replication. It is likely that the 54 CTDGs identified here include novel HIV-1 co-factors. The presence of CTDGs in the protein space that was available for HIV-1 to sample during its evolution and acquisition of Tat function may provide an explanation for why CTDGs are enriched in viral co-factors.
HIV-1在CD4(+) T细胞和巨噬细胞中介导其复制周期依赖于细胞辅助因子,这两种主要细胞类型是该病毒在体内感染的对象。一个关键的辅助因子是细胞周期蛋白T1,它是一种称为P-TEFb的通用RNA聚合酶II延伸因子的亚基。细胞周期蛋白T1被病毒Tat蛋白直接靶向以激活前病毒转录。当静息CD4(+) T细胞被激活时以及在巨噬细胞分化或激活过程中,细胞周期蛋白T1会上调,而这些条件对于高水平的HIV-1复制也是必需的。由于细胞周期蛋白T1是转录因子的一个亚基,这些细胞中细胞周期蛋白T1的上调会导致细胞基因的诱导,其中一些可能是HIV-1辅助因子。通过使用细胞周期蛋白T1的短发夹RNA(shRNA)消耗和转录谱分析,我们鉴定出54种细胞mRNA,它们在活化的CD4(+) T Jurkat T细胞以及人单核细胞系MM6细胞的分化和激活过程中,其诱导似乎依赖于细胞周期蛋白T1。这些细胞周期蛋白T1依赖性基因(CTDG)的启动子在两个转录因子结合位点SREBP1和ARP1中过度富集。值得注意的是,据报道这些CTDG中有10个参与HIV-1复制,与随机生成的54个基因列表相比,此类基因的过度富集非常显著(p值<0.00021)。这里鉴定出的两个CTDG(CDK11和酪蛋白激酶1γ1)的小干扰RNA(siRNA)消耗和显性负性蛋白实验结果表明,这些基因直接或间接参与HIV-1复制。这里鉴定出的54个CTDG很可能包括新的HIV-1辅助因子。在HIV-1进化和获得Tat功能期间可用于其采样的蛋白质空间中存在CTDG,这可能解释了为什么CTDG在病毒辅助因子中富集。