Elongin A regulates transcription in vivo through enhanced RNA polymerase processivity
| Autor: | Brian David Dynlacht, Yating Wang, M. Behfar Ardehali, Liming Hou, Robert E. Kingston |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
0301 basic medicine
FPKM fragments per kilobase per million mapped reads Transcription Elongation Genetic Elongin Enhancer RNAs RNA polymerase II Elongin complex Biochemistry CUT&RUN cleavage under targets and release using nuclease 4sU 4-thiouridine chemistry.chemical_compound eRNA enhancer RNA Transcription (biology) RNA polymerase Serine Transcriptional regulation Phosphorylation RNA Small Interfering Ser2 serine 2 biology Chemistry 4sU-seq 4sU-labeled RNA sequencing Chromatin Cell biology ChIP chromatin immunoprecipitation Enhancer Elements Genetic transcription Signal Transduction Research Article 03 medical and health sciences Cell Line Tumor Elongin A Humans RNA Messenger transcription elongation Enhancer Molecular Biology 030102 biochemistry & molecular biology Sequence Analysis RNA TESs transcript end sites RNAPII RNA polymerase II ES embryonic stem Computational Biology Epithelial Cells Cell Biology 030104 developmental biology Gene Expression Regulation biology.protein TSS transcription start site DRB 5 6-dichloro-1-beta-D-ribofuranosylbenzimidazole enhancer 5 6-Dichloro-1-beta-D-ribofuranosylbenzimidazole |
| Zdroj: | The Journal of Biological Chemistry |
| ISSN: | 0021-9258 |
| DOI: | 10.1074/jbc.ra120.015876 |
| Popis: | Elongin is an RNA polymerase II (RNAPII)-associated factor that has been shown to stimulate transcriptional elongation in vitro. The Elongin complex is thought to be required for transcriptional induction in response to cellular stimuli and to ubiquitinate RNAPII in response to DNA damage. Yet, the impact of the Elongin complex on transcription in vivo has not been well studied. Here, we performed comprehensive studies of the role of Elongin A, the largest subunit of the Elongin complex, on RNAPII transcription genome-wide. Our results suggest that Elongin A localizes to actively transcribed regions and potential enhancers, and the level of recruitment correlated with transcription levels. We also identified a large group of factors involved in transcription as Elongin A–associated factors. In addition, we found that loss of Elongin A leads to dramatically reduced levels of serine2-phosphorylated, but not total, RNAPII, and cells depleted of Elongin A show stronger promoter RNAPII pausing, suggesting that Elongin A may be involved in the release of paused RNAPII. Our RNA-seq studies suggest that loss of Elongin A did not alter global transcription, and unlike prior in vitro studies, we did not observe a dramatic impact on RNAPII elongation rates in our cell-based nascent RNA-seq experiments upon Elongin A depletion. Taken together, our studies provide the first comprehensive analysis of the role of Elongin A in regulating transcription in vivo. Our studies also revealed that unlike prior in vitro findings, depletion of Elongin A has little impact on global transcription profiles and transcription elongation in vivo. |
| Databáze: | OpenAIRE |
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