Bhandare, Pranjali and Narain, Ashwin and Hofstetter, Julia and Rummel, Teresa and Wenzel, Julia and Schuelein-Voelk, Christina and Lamer, Stephanie and Eilers, Ursula and Schlosser, Andreas and Eilers, Martin and Erhard, Florian and Wolf, Elmar (2024) Phenotypic screens identify SCAF1 as critical activator of RNAPII elongation and global transcription. NUCLEIC ACIDS RESEARCH, 53 (4). ISSN 0305-1048, 1362-4962
Full text not available from this repository. (Request a copy)Abstract
Transcripts produced by RNA polymerase II (RNAPII) are fundamental for cellular responses to environmental changes. It is therefore no surprise that there exist multiple avenues for the regulation of this process. To explore the regulation mediated by RNAPII-interacting proteins, we used a small interfering RNA (siRNA)-based screen to systematically evaluate their influence on RNA synthesis. We identified several proteins that strongly affected RNAPII activity. We evaluated one of the top hits, SCAF1 (SR-related C-terminal domain-associated factor 1), using an auxin-inducible degradation system and sequencing approaches. In agreement with our screen results, acute depletion of SCAF1 decreased RNA synthesis, and showed an increase of Serine-2 phosphorylated-RNAPII (pS2-RNAPII). We found that the accumulation of pS2-RNAPII within the gene body occurred at GC-rich regions and was indicative of stalled RNAPII complexes. The accumulation of stalled RNAPII complexes was accompanied by reduced recruitment of initiating RNAPII, explaining the observed global decrease in transcriptional output. Furthermore, upon SCAF1 depletion, RNAPII complexes showed increased association with components of the proteasomal-degradation machinery. We concluded that in cells lacking SCAF1, RNAPII undergoes a rather interrupted passage, resulting in intervention by the proteasomal-degradation machinery to clear stalled RNAPII. While cells survive the compromised transcription caused by absence of SCAF1, further inhibition of proteasomal-degradation machinery is synthetically lethal.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | PRE-MESSENGER-RNA; POLYMERASE-II CTD; C-TERMINAL DOMAIN; DNA-DAMAGE; HUMAN SPLICEOSOME; GENE-EXPRESSION; PROTEIN; DEGRADATION; PHOSPHORYLATION; UBIQUITYLATION |
| Subjects: | 000 Computer science, information & general works > 004 Computer science 600 Technology > 610 Medical sciences Medicine |
| Divisions: | Informatics and Data Science > Department Computational Life Science > Chair of Computational Immunology (Prof. Dr. Florian Erhard) |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 27 Jan 2026 09:58 |
| Last Modified: | 27 Jan 2026 09:58 |
| URI: | https://pred.uni-regensburg.de/id/eprint/65222 |
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