MAF1 represses CDKN1A through a Pol III-dependent mechanism

Elife. 2015 Jun 12:4:e06283. doi: 10.7554/eLife.06283.

Abstract

MAF1 represses Pol III-mediated transcription by interfering with TFIIIB and Pol III. Herein, we found that MAF1 knockdown induced CDKN1A transcription and chromatin looping concurrently with Pol III recruitment. Simultaneous knockdown of MAF1 with Pol III or BRF1 (subunit of TFIIIB) diminished the activation and looping effect, which indicates that recruiting Pol III was required for activation of Pol II-mediated transcription and chromatin looping. Chromatin-immunoprecipitation analysis after MAF1 knockdown indicated enhanced binding of Pol III and BRF1, as well as of CFP1, p300, and PCAF, which are factors that mediate active histone marks, along with the binding of TATA binding protein (TBP) and POLR2E to the CDKN1A promoter. Simultaneous knockdown with Pol III abolished these regulatory events. Similar results were obtained for GDF15. Our results reveal a novel mechanism by which MAF1 and Pol III regulate the activity of a protein-coding gene transcribed by Pol II.

Keywords: CDKN1A; MAF1; Pol III; biochemistry; cell biology; human.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Line
  • Chromatin Immunoprecipitation
  • Cyclin-Dependent Kinase Inhibitor p21 / biosynthesis*
  • DNA / metabolism
  • Gene Expression Regulation*
  • Gene Knockdown Techniques
  • Humans
  • Promoter Regions, Genetic
  • Protein Binding
  • RNA Polymerase III / metabolism*
  • Repressor Proteins / genetics
  • Repressor Proteins / metabolism*

Substances

  • CDKN1A protein, human
  • Cyclin-Dependent Kinase Inhibitor p21
  • MAF1 protein, human
  • Repressor Proteins
  • DNA
  • RNA Polymerase III

Grants and funding

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.