MAF1 represses CDKN1A through a Pol III-dependent mechanism

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...

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Published in:eLife 2015-06, Vol.4, p.e06283-e06283
Main Authors: Lee, Yu-Ling, Li, Yuan-Ching, Su, Chia-Hsin, Chiao, Chun-Hui, Lin, I-Hsuan, Hsu, Ming-Ta
Format: Article
Language:English
Subjects:
Biochemistry
CDKN1A
Cell Biology
Cell Line
Chromatin Immunoprecipitation
Cyclin-Dependent Kinase Inhibitor p21 - biosynthesis
DNA - metabolism
Gene Expression Regulation
Gene Knockdown Techniques
Humans
MAF1
Pol III
Promoter Regions, Genetic
Protein Binding
Repressor Proteins - genetics
Repressor Proteins - metabolism
RNA Polymerase III - metabolism
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Summary: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.
ISSN:2050-084X
2050-084X
DOI:10.7554/eLife.06283