Arginine methylation of the DDX 5 helicase RGG / RG motif by PRMT 5 regulates resolution of RNA:DNA hybrids
| Autor: | Jean-Yves Masson, Stéphane Richard, Martin Karam, Zhenbao Yu, Franciele F. Busatto, Yan Coulombe, Sofiane Y Mersaoui |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
Protein-Arginine N-Methyltransferases
RNA helicase Amino Acid Motifs RNA polymerase II Biology Arginine Methylation Article General Biochemistry Genetics and Molecular Biology Cell Line DEAD-box RNA Helicases 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Arginine methylation Transcription (biology) RGG/RG motif Exoribonuclease DDX5 Humans XRN2 Molecular Biology 030304 developmental biology 0303 health sciences General Immunology and Microbiology General Neuroscience DNA Replication Repair & Recombination Post-translational Modifications Proteolysis & Proteomics Helicase RNA DNA Articles RNA Biology RNA Helicase A Cell biology HEK293 Cells chemistry Exoribonucleases biology.protein RNA Polymerase II 030217 neurology & neurosurgery |
| Zdroj: | The EMBO Journal |
| ISSN: | 1460-2075 0261-4189 |
| Popis: | Aberrant transcription‐associated RNA:DNA hybrid (R‐loop) formation often causes catastrophic conflicts during replication, resulting in DNA double‐strand breaks and genomic instability. Preventing such conflicts requires hybrid dissolution by helicases and/or RNase H. Little is known about how such helicases are regulated. Herein, we identify DDX5, an RGG/RG motif‐containing DEAD‐box family RNA helicase, as crucial player in R‐loop resolution. In vitro, recombinant DDX5 resolves R‐loops in an ATP‐dependent manner, leading to R‐loop degradation by the XRN2 exoribonuclease. DDX5‐deficient cells accumulate R‐loops at loci with propensity to form such structures based on RNA:DNA immunoprecipitation (DRIP)‐qPCR, causing spontaneous DNA double‐strand breaks and hypersensitivity to replication stress. DDX5 associates with XRN2 and resolves R‐loops at transcriptional termination regions downstream of poly(A) sites, to facilitate RNA polymerase II release associated with transcriptional termination. Protein arginine methyltransferase 5 (PRMT5) binds and methylates DDX5 at its RGG/RG motif. This motif is required for DDX5 interaction with XRN2 and repression of cellular R‐loops, but not essential for DDX5 helicase enzymatic activity. PRMT5‐deficient cells accumulate R‐loops, resulting in increased formation of γH2AX foci. Our findings exemplify a mechanism by which an RNA helicase is modulated by arginine methylation to resolve R‐loops, and its potential role in regulating transcription. |
| Databáze: | OpenAIRE |
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