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Author Notes:

Ruichuan Chen. Tel: +86 592 2184533; Fax: +86 592 2181015; Email: chenrc@xmu.edu.cn

Runzhong Liu. Tel: +86 592 2184533; Fax: +86 592 2181015; Email: liurz@xmu.edu.cn

Lin-Feng Chen. Tel: +1 217 333 7764; Fax: +1 217 244 5858; Email: lfchen@life.illinois.edu

R.C, and R.L. designed the project. X.L. and X.Z. performed most of the experiments. Y.L., and K.Z. designed and performed MS analysis. M.L., X.L., B.Y., H.Y. and Y.W. designed and performed ChIP-Seq, ChIP-silver staining and ChIP-qPCR analysis. Y.W. and M.R. performed qRT-PCR analysis. Y.W., K.Z., Y.H., Y.W., Y.C. and M.C. performed cDNA cloning and shRNA preparation. Y.L. and X.L. carried out kinase assay. R.C. and L.C. write the manuscript. All authors commented on the manuscript.

The authors thank Analysis and Testing Center of School of Life Sciences of Xiamen University for MS analysis and Dr Zhong, Chuangqi for help of the calculation of MS data.

Conflict of interest statement. None declared.

Subjects:

Research Funding:

National Natural Science Foundation of China [81361120386, 31570751, 31270809 and 30930046 to R.C., 31501096 to M.L., 81171192 to R.L.]; 973 program [2013CB917802 to R.C.]; NIH-CA179511 [to L.F.C.]; Fundamental Research Funds for the Central Universities [20720150053 to M.L.]; NSFC for Fostering Talents in Basic Research [J1310027 to Y.C]. Funding for open access charge: National Natural Science Foundation of China [81361120386, 31570751, 31270809 and 30930046 to R.C., 31501096 to M.L., 81171192 to R.L.]; 973 program [2013CB917802 to R.C.]; NIH-CA179511 [to L.F.C.]; Fundamental Research Funds for the Central Universities [20720150053 to M.L.]; NSFC for Fostering Talents in Basic Research [J1310027 to Y.C].

Keywords:

  • Acetamides
  • Cell Cycle Proteins
  • HCT116 Cells
  • HeLa Cells
  • Humans
  • Models, Biological
  • Nuclear Proteins
  • Phosphorylation
  • Positive Transcriptional Elongation Factor B
  • Promoter Regions, Genetic
  • RNA Polymerase II
  • RNA, Small Interfering
  • Transcription Elongation, Genetic
  • Transcription Factors
  • Transcriptional Elongation Factors

Multiple P-TEFbs cooperatively regulate the release of promoter-proximally paused RNA polymerase II.

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Journal Title:

Nucleic Acids Res

Volume:

Volume 44, Number 14

Publisher:

, Pages 6853-6867

Type of Work:

Article | Final Publisher PDF

Abstract:

The association of DSIF and NELF with initiated RNA Polymerase II (Pol II) is the general mechanism for inducing promoter-proximal pausing of Pol II. However, it remains largely unclear how the paused Pol II is released in response to stimulation. Here, we show that the release of the paused Pol II is cooperatively regulated by multiple P-TEFbs which are recruited by bromodomain-containing protein Brd4 and super elongation complex (SEC) via different recruitment mechanisms. Upon stimulation, Brd4 recruits P-TEFb to Spt5/DSIF via a recruitment pathway consisting of Med1, Med23 and Tat-SF1, whereas SEC recruits P-TEFb to NELF-A and NELF-E via Paf1c and Med26, respectively. P-TEFb-mediated phosphorylation of Spt5, NELF-A and NELF-E results in the dissociation of NELF from Pol II, thereby transiting transcription from pausing to elongation. Additionally, we demonstrate that P-TEFb-mediated Ser2 phosphorylation of Pol II is dispensable for pause release. Therefore, our studies reveal a co-regulatory mechanism of Brd4 and SEC in modulating the transcriptional pause release by recruiting multiple P-TEFbs via a Mediator- and Paf1c-coordinated recruitment network.

Copyright information:

© The Author(s) 2016. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access work distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International License (https://creativecommons.org/licenses/by-nc/4.0/rdf).
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