Recruitment of TFIIH to the HIV LTR is a rate-limiting step in the emergence of HIV from latency.
Latently infected cells rapidly initiate HIV transcription after exposure to signals that induce NF-kappaB. To investigate the role of TFIIH during HIV reactivation in vivo, we developed a population of Jurkat cells containing integrated, but transcriptionally silent, HIV proviruses. Surprisingly, the HIV promoter in unactivated Jurkat T cells is partially occupied and carries Mediator containing the CDK8 repressive module, TFIID and RNAP II that is hypophosphorylated and confined to the promoter region. Significantly, the promoter is devoid of TFIIH. Upon stimulation of the cells by TNF-alpha, NF-kappaB and TFIIH are rapidly recruited to the promoter together with additional Mediator and RNAP II, but CDK8 is lost. Detailed time courses show that the levels of TFIIH at the promoter fluctuate in parallel with NF-kappaB recruitment to the promoter. Similarly, recombinant p65 activates HIV transcription in vitro and stimulates phosphorylation of the RNAP II CTD by the CDK7 kinase module of TFIIH. We conclude that the recruitment and activation of TFIIH represents a rate-limiting step for the emergence of HIV from latency.
Kim
Young Kyeung
YK
Department of Molecular Biology and Microbiology, School of Medicine, Case Western Reserve University, Cleveland, OH 44106, USA.
Bourgeois
Cyril F
CF
Pearson
Richard
R
Tyagi
Mudit
M
West
Michelle J
MJ
Wong
Julian
J
Wu
Shwu-Yuan
SY
Chiang
Cheng-Ming
CM
Karn
Jonathan
J
D015967:Gene Expression Regulation, Viral; D006678:HIV / Q000235:genetics* / Q000378:metabolism / Q000502:physiology; D016325:HIV Long Terminal Repeat; D006801:Humans; D019169:Jurkat Cells; D008954:Models, Biological; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism; D010766:Phosphorylation; D011401:Promoter Regions, Genetic; D012319:RNA Polymerase II / Q000235:genetics / Q000378:metabolism; D013997:Time Factors; D051758:Transcription Factor TFIIH / Q000235:genetics / Q000378:metabolism*; D014158:Transcription, Genetic; D017735:Virus Latency / Q000235:genetics*
D016328:NF-kappa B; D051758:Transcription Factor TFIIH; D012319:RNA Polymerase II