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PMID: 23589332 Published · ppublish English Journal Article Research Support, N.I.H., Intramural

BRD4 coordinates recruitment of pause release factor P-TEFb and the pausing complex NELF/DSIF to regulate transcription elongation of interferon-stimulated genes.

Molecular and cellular biology ·Vol. 33 ·No. 12 ·2013-06-00 ·Pages 2497-507

Patel MC, Debrosse M, Smith M, Dey A, Huynh W, Sarai N, Heightman TD, Tamura T, Ozato K

Abstract

RNA polymerase II (Pol II) and the pausing complex, NELF and DSIF, are detected near the transcription start site (TSS) of many active and silent genes. Active transcription starts when the pause release factor P-TEFb is recruited to initiate productive elongation. However, the mechanism of P-TEFb recruitment and regulation of NELF/DSIF during transcription is not fully understood. We investigated this question in interferon (IFN)-stimulated transcription, focusing on BRD4, a BET family protein that interacts with P-TEFb. Besides P-TEFb, BRD4 binds to acetylated histones through the bromodomain. We found that BRD4 and P-TEFb, although not present prior to IFN treatment, were robustly recruited to IFN-stimulated genes (ISGs) after stimulation. Likewise, NELF and DSIF prior to stimulation were hardly detectable on ISGs, which were strongly recruited after IFN treatment. A shRNA-based knockdown assay of NELF revealed that it negatively regulates the passage of Pol II and DSIF across the ISGs during elongation, reducing total ISG transcript output. Analyses with a BRD4 small-molecule inhibitor showed that IFN-induced recruitment of P-TEFb and NELF/DSIF was under the control of BRD4. We suggest a model where BRD4 coordinates both positive and negative regulation of ISG elongation.

MeSH Terms
3T3 Cells Animals Azepines/pharmacology Cell Line Cyclin-Dependent Kinase 9/metabolism Interferon-beta/metabolism Mice Nuclear Proteins/genetics,metabolism Positive Transcriptional Elongation Factor B/metabolism Promoter Regions, Genetic RNA Interference RNA Polymerase II RNA, Small Interfering Transcription Factors/genetics,metabolism Transcription Initiation Site Transcription, Genetic Triazoles/pharmacology
Chemicals
(+)-JQ1 compound Azepines Brd4 protein, mouse DSIF protein, mouse NELF protein, mouse Nuclear Proteins RNA, Small Interfering Transcription Factors Triazoles Interferon-beta Positive Transcriptional Elongation Factor B Cdk9 protein, mouse Cyclin-Dependent Kinase 9 RNA Polymerase II
Authors & Affiliations
9 authors, click to expand affiliations / ORCID
Patel Mira C
Program in Genomics of Differentiation, NICHD, National Institutes of Health, Bethesda, Maryland, USA.
Debrosse Maxime
Smith Matthew
Dey Anup
Huynh Walter
Sarai Naoyuki
Heightman Tom D
Tamura Tomohiko
Ozato Keiko
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
1098-5549
DOI
10.1128/MCB.01180-12
Published
2013-06-00
Epub
2013-00-15
Pages
2497-507
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC3700095
Subset
IM
Grants
Intramural NIH HHS · United States
External Links
PubMed source DOI: 10.1128/MCB.01180-12 PMC Full Text View journal details
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