Barton Kirston, Margolis David
Department of Medicine, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599-7042, USA.
AIDS Res Hum Retroviruses. 2013 Feb;29(2):289-98. doi: 10.1089/AID.2012.0227. Epub 2012 Sep 4.
Quiescent HIV-1 infection of resting CD4(+) T cells is an obstacle to eradication of HIV-1 infection. These reservoirs are maintained, in part, by repressive complexes that bind to the HIV-1 long terminal repeat (LTR) and recruit histone deacetylases (HDACs). cMyc and YY1 are two transcription factors that are recruited as part of well-described, distinct complexes to the HIV-1 LTR and in turn recruit HDACs. In prior studies, depletion of single factors that recruit HDAC1 in various cell lines was sufficient to upregulate LTR activity. We used short hairpin RNAs (shRNAs) to test the effect of targeted disruption of a single transcription factor on quiescent proviruses in T cell lines. In this study, we found that depletion of YY1 significantly increases mRNA and protein expression from the HIV-1 promoter in some contexts, but does not affect HDAC1, HDAC2, HDAC3, or acetylated histone 3 occupancy of the HIV-1 LTR. Conversely, depletion of cMyc or cMyc and YY1 does not significantly alter the level of transcription from the LTR or affect recruitment of HDACs to the HIV-1 LTR. Furthermore, global inhibition of HDACs with the HDAC inhibitor suberoylanilide hydroxamic acid (SAHA) enhanced the increase in LTR transcription in cells that were depleted of YY1.These findings show that despite prior isolated findings, redundancy in repressors of HIV-1 LTR expression will require selective targeting of multiple restrictive mechanisms to comprehensively induce the escape of quiescent proviruses from latency.
静息CD4(+) T细胞的潜伏性HIV-1感染是根除HIV-1感染的一个障碍。这些病毒库部分由与HIV-1长末端重复序列(LTR)结合并募集组蛋白脱乙酰酶(HDAC)的抑制复合物维持。cMyc和YY1是两种转录因子,它们作为已充分描述的不同复合物的一部分被募集到HIV-1 LTR,进而募集HDAC。在先前的研究中,在各种细胞系中耗尽募集HDAC1的单一因子足以上调LTR活性。我们使用短发夹RNA(shRNA)来测试靶向破坏单个转录因子对T细胞系中潜伏前病毒的影响。在本研究中,我们发现,在某些情况下,YY1的耗尽显著增加了HIV-1启动子的mRNA和蛋白质表达,但不影响HDAC1、HDAC2、HDAC3或HIV-1 LTR上乙酰化组蛋白3的占有率。相反,cMyc或cMyc和YY1的耗尽不会显著改变LTR的转录水平,也不会影响HDAC募集到HIV-1 LTR。此外,用HDAC抑制剂辛二酰苯胺异羟肟酸(SAHA)对HDAC进行全局抑制增强了YY1耗尽细胞中LTR转录的增加。这些发现表明,尽管有先前的孤立发现,但HIV-1 LTR表达抑制因子的冗余将需要对多种限制机制进行选择性靶向,以全面诱导潜伏前病毒从潜伏状态中逃逸。