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| Status |
Public on Jun 10, 2015 |
| Title |
MAF1 Represses CDKN1A through a Pol III-Dependent Mechanism |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by array
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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. ChIP 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 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.
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| Overall design |
Knockdown assay was performed using siRNA obtained from MISSION®RNA (Sigma). Inhibition of expression of Pol III (SASI_Hs01_00046568) and MAF1 (SASI_Hs01_00135954) was achieved by transfection with LipofectamineTM RNAiMax (Invitrogen) according to the manufacturer’s protocol. MISSION® siRNA Universal Negative Control (Sigma) was used as knockdown control. Cells were transfected in serum-free medium. After 8 h, the siRNA containing medium was replaced with complete medium.
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| Contributor(s) |
Lee Y, Li Y |
| Citation(s) |
26067234 |
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| Submission date |
Nov 13, 2012 |
| Last update date |
Mar 25, 2019 |
| Contact name |
Ming-Ta Hsu |
| E-mail(s) |
kentkenpony@hotmail.com
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| Organization name |
National Yang Ming University
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| Street address |
No.155, Sec.2, Linong Street
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| City |
Taipei |
| ZIP/Postal code |
112 |
| Country |
Taiwan |
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| Platforms (1) |
| GPL570 |
[HG-U133_Plus_2] Affymetrix Human Genome U133 Plus 2.0 Array |
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| Samples (3) |
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| Relations |
| BioProject |
PRJNA179433 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE42239_RAW.tar |
14.7 Mb |
(http)(custom) |
TAR (of CEL, CHP) |
| GSE42239_fold_change.txt.gz |
3.7 Mb |
(ftp)(http) |
TXT |
| Processed data included within Sample table |
| Processed data provided as supplementary file |
| Processed data are available on Series record |
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