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| Status |
Public on Aug 09, 2022 |
| Title |
SPT6 functions in transcriptional pause-release via PAF1C recruitment [RNA-seq] |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
In vitro studies identified various factors including P-TEFb, SEC, SPT6, PAF1, DSIF, and NELF functioning at different stages of transcription elongation driven by RNA polymerase II (RNA Pol II). What remains unclear is how these factors cooperatively regulate pause/release and productive elongation in the context of living cells. Using an acute protein-depletion approach, we report that SPT6 depletion results in release of paused RNA Pol II. Short genes demonstrate a prominent release and a subsequent increase in mature transcripts, whereas long genes fail to yield mature transcripts due to a loss of processivity. Unexpectedly, the recruitment of PAF1 complex (PAF1C) to RNA Pol II fails upon SPT6 depletion, leading to the release of NELF-bound RNA Pol II into the gene bodies. Furthermore, SPT6 depletion impairs heat shock-induced pausing, pointing to a role for SPT6 in regulating RNA Pol II pause-release through PAF1C recruitment and NELF removal during the early elongation.
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| Overall design |
Examination of SPT6 functions in human cells using acute and rapid depletion system and high-throughput sequencing (ChIP-seq, RNA-seq, PRO-seq).
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| Contributor(s) |
Aoi Y, Shilatifard A |
| Citation(s) |
35973425 |
| |
| Submission date |
May 04, 2022 |
| Last update date |
Nov 09, 2022 |
| Contact name |
Yuki Aoi |
| E-mail(s) |
yuki.aoi@northwestern.edu
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| Organization name |
Northwestern University
|
| Department |
Biochemistry & Molecular Genetics
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| Lab |
Ali Shilatifard
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| Street address |
303 E Superior St. SQBRC 7-300
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| City |
Chicago |
| State/province |
IL |
| ZIP/Postal code |
60611 |
| Country |
USA |
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| Platforms (1) |
| GPL24676 |
Illumina NovaSeq 6000 (Homo sapiens) |
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| Samples (12)
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| This SubSeries is part of SuperSeries: |
| GSE202190 |
SPT6 functions in transcriptional pause-release via PAF1C recruitment |
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| Relations |
| BioProject |
PRJNA834960 |
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