TFIID Enables RNA Polymerase II Promoter-Proximal Pausing

Charli B Fant, Cecilia B Levandowski, Kapil Gupta, Zachary L Maas, John Moir, Jonathan D Rubin, Andrew Sawyer, Meagan N Esbin, Jenna K Rimel, Olivia Luyties, Michael T Marr, Imre Berger, Robin D Dowell, Dylan J Taatjes*

*Corresponding author for this work

Research output: Contribution to journalEditorial (Academic Journal)peer-review

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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
Original languageEnglish
Pages (from-to)785-793.e8
Number of pages18
JournalMolecular Cell
Volume78
Issue number4
Early online date30 Mar 2020
DOIs
Publication statusPublished - 21 May 2020

Keywords

  • TFIID
  • TRIM-Away
  • TAF1
  • RNA polymerase II
  • pausing
  • NELF
  • DSIF
  • TBP
  • P-TEFb
  • PRO-seq

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