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| Status |
Public on Dec 20, 2017 |
| Title |
Association with Aurora-A controls N-MYC-dependent promoter escape and pause release of RNA polymerase II during the cell cycle |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by high throughput sequencing
Genome binding/occupancy profiling by high throughput sequencing
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| Summary |
MYC proteins bind globally to active promoters and promote transcriptional elongation by RNA polymerase II (RNAPII). To identify effector proteins that mediate this function, we performed mass spectrometry on N-MYC complexes in neuroblastoma cells. The analysis shows that N-MYC forms complexes with TFIIIC, TOP2A and RAD21, a subunit of cohesin. N-MYC and TFIIIC bind to overlapping sites in thousands of RNAPII promoters and intergenic regions. TFIIIC promotes association of RAD21 with N-MYC target sites and is required for N-MYC-dependent promoter escape and pause release of RNAPII. Aurora-A competes with binding of TFIIIC and RAD21 to N-MYC in vitro and antagonizes association of TOP2A, TFIIIC and RAD21 with N-MYC during S-phase, blocking N-MYC-dependent release of RNAPII from the promoter. Inhibition of Aurora-A in S-phase restores RAD21 and TFIIIC binding to chromatin and partially restores N-MYC-dependent transcriptional elongation. We propose that complex formation with Aurora-A controls N-MYC function during the cell cycle.
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| Overall design |
DNA-binding of N-MYC and TFIIIC upon inhibition of Aurora-A, RNAPII occupancy after shRNA-mediated depletion of TFIIIC5 or in S-phase arrested SH-EP-NMYCER cells and gene expression changes after depletion of RAD21, Aurora-A and TFIIIC5 or activation of NMYC.
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| Contributor(s) |
Büchel G, Carstensen A, Mak K, Roeschert I, Leen E, Sumara O, Hofstetter J, Herold S, Kalb J, Baluapuri A, Poon E, Kwok C, Chesler L, Maric HM, Rickman DS, Wolf E, Bayliss R, Walz S, Eilers M |
| Citation(s) |
29262328 |
| Submission date |
Mar 07, 2016 |
| Last update date |
Jul 25, 2021 |
| Contact name |
Martin Eilers |
| Organization name |
University of Wuerzburg
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| Department |
Chair for Biochemistry and Molecular Biology
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| Lab |
Martin Eilers
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| Street address |
Am Hubland
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| City |
Wuerzburg |
| ZIP/Postal code |
97074 |
| Country |
Germany |
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| Platforms (2) |
| GPL10999 |
Illumina Genome Analyzer IIx (Homo sapiens) |
| GPL18573 |
Illumina NextSeq 500 (Homo sapiens) |
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| Samples (50)
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| Relations |
| BioProject |
PRJNA314499 |
| SRA |
SRP071219 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE78957_CD532_4h_vs_DMSO.txt.gz |
958.1 Kb |
(ftp)(http) |
TXT |
| GSE78957_CD532_8h_vs_DMSO.txt.gz |
984.5 Kb |
(ftp)(http) |
TXT |
| GSE78957_RAW.tar |
2.5 Gb |
(http)(custom) |
TAR (of BEDGRAPH, WIG) |
| GSE78957_RNAseq_raw_count_matrix.txt.gz |
980.9 Kb |
(ftp)(http) |
TXT |
| GSE78957_SHEP_NMYCER_plusOHT_vs_minusOHT.txt.gz |
672.1 Kb |
(ftp)(http) |
TXT |
| GSE78957_siAURKA_vs_siCTR.txt.gz |
1012.6 Kb |
(ftp)(http) |
TXT |
| GSE78957_siRAD21_vs_siCTR.txt.gz |
973.6 Kb |
(ftp)(http) |
TXT |
| GSE78957_siTF3C5_vs_siCTR.txt.gz |
865.6 Kb |
(ftp)(http) |
TXT |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data are available on Series record |
| Processed data provided as supplementary file |
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