Benkirane M, Chun R F, Xiao H, Ogryzko V V, Howard B H, Nakatani Y, Jeang K T
Molecular Virology Section, Laboratory of Molecular Microbiology, NIAID, National Institutes of Health, Bethesda, Maryland 20892, USA.
J Biol Chem. 1998 Sep 18;273(38):24898-905. doi: 10.1074/jbc.273.38.24898.
A unique aspect of the retrovirus life cycle is the obligatory integration of the provirus into host cell chromosomes. Unlike viruses that do not integrate, retroviruses must conserve an ability to activate transcription from a chromatin context. Human immunodeficiency virus (HIV)-1 encodes an unusual and an unusually potent transcriptional transactivator, Tat, which binds to a nascent viral leader RNA, TAR. The action of Tat has been well studied in various reductive model systems; however, the physiological mechanism through which Tat gains access to chromatin-associated proviral long terminal repeats (LTRs) is not understood. We show here that a nuclear histone acetyltransferase activity associates with Tat. Intracellularly, we found that Tat forms a ternary complex with p300 and P/CAF, two histone acetyltransferases (HATs). A murine cell defect in Tat transactivation of the HIV-1 LTR was linked to the reduced abundance of p300 and P/CAF. Thus, overexpression of p300 and P/CAF reconstituted Tat transactivation of the HIV-1 LTR in NIH3T3 cells to a level similar to that observed for human cells. By using transdominant p300 or P/CAF mutants that lack enzymatic activity, we delineated a requirement for the HAT component from the latter but not the former in Tat function. Finally, we observed that Tat-associated HAT is preferentially important for transactivation of integrated, but not unintegrated, HIV-1 LTR.
逆转录病毒生命周期的一个独特方面是前病毒必须整合到宿主细胞染色体中。与不进行整合的病毒不同,逆转录病毒必须保留从染色质环境激活转录的能力。人类免疫缺陷病毒1型(HIV-1)编码一种不同寻常且异常强大的转录反式激活因子Tat,它能与新生的病毒前导RNA(TAR)结合。Tat的作用已在各种简化模型系统中得到充分研究;然而,Tat进入与染色质相关的前病毒长末端重复序列(LTRs)的生理机制尚不清楚。我们在此表明,一种核组蛋白乙酰转移酶活性与Tat相关。在细胞内,我们发现Tat与两种组蛋白乙酰转移酶(HATs)p300和P/CAF形成三元复合物。HIV-1 LTR的Tat反式激活在鼠细胞中的缺陷与p300和P/CAF丰度降低有关。因此,p300和P/CAF的过表达将NIH3T3细胞中HIV-1 LTR的Tat反式激活恢复到与人类细胞中观察到的水平相似。通过使用缺乏酶活性的显性负性p300或P/CAF突变体,我们确定了Tat功能对后者而非前者的HAT成分的需求。最后,我们观察到与Tat相关的HAT对整合型而非未整合型HIV-1 LTR的反式激活尤为重要。