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TFIID Enables RNA Polymerase II Promoter-Proximal Pausing.

Authors
  • Fant, Charli B1
  • Levandowski, Cecilia B1
  • Gupta, Kapil2
  • Maas, Zachary L1
  • Moir, John1
  • Rubin, Jonathan D1
  • Sawyer, Andrew3
  • Esbin, Meagan N1
  • Rimel, Jenna K1
  • Luyties, Olivia1
  • Marr, Michael T3
  • Berger, Imre2
  • Dowell, Robin D4
  • Taatjes, Dylan J5
  • 1 Department of Biochemistry, University of Colorado, Boulder, CO, USA.
  • 2 School of Biochemistry, Bristol Research Centre for Synthetic Biology, University of Bristol, Bristol, UK.
  • 3 Department of Biology, Brandeis University, Waltham, MA, USA.
  • 4 Department of Molecular, Cellular, and Developmental Biology, University of Colorado, Boulder, CO, USA; BioFrontiers Institute, University of Colorado, Boulder, CO, USA.
  • 5 Department of Biochemistry, University of Colorado, Boulder, CO, USA. Electronic address: [email protected]
Type
Published Article
Journal
Molecular cell
Publication Date
May 21, 2020
Volume
78
Issue
4
Identifiers
DOI: 10.1016/j.molcel.2020.03.008
PMID: 32229306
Source
Medline
Keywords
Language
English
License
Unknown

Abstract

RNA polymerase II (RNAPII) transcription is governed by the pre-initiation complex (PIC), which contains TFIIA, TFIIB, TFIID, TFIIE, TFIIF, TFIIH, RNAPII, and Mediator. After initiation, RNAPII enzymes pause after transcribing less than 100 bases; precisely how RNAPII pausing is enforced and regulated remains unclear. To address specific mechanistic questions, we reconstituted human RNAPII promoter-proximal pausing in vitro, entirely with purified factors (no extracts). As expected, NELF and DSIF increased pausing, and P-TEFb promoted pause release. Unexpectedly, the PIC alone was sufficient to reconstitute pausing, suggesting RNAPII pausing is an inherent PIC function. In agreement, pausing was lost upon replacement of the TFIID complex with TATA-binding protein (TBP), and PRO-seq experiments revealed widespread disruption of RNAPII pausing upon acute depletion (t = 60 min) of TFIID subunits in human or Drosophila cells. These results establish a TFIID requirement for RNAPII pausing and suggest pause regulatory factors may function directly or indirectly through TFIID. Copyright © 2020 Elsevier Inc. All rights reserved.

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