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EMBO J. 2019 Aug 1;38(15):e100986. doi: 10.15252/embj.2018100986. Epub 2019 Jun 21.

Arginine methylation of the DDX5 helicase RGG/RG motif by PRMT5 regulates resolution of RNA:DNA hybrids.

Author information

1
Departments of Oncology and Medicine, Segal Cancer Center, Lady Davis Institute for Medical Research, McGill University, Montréal, QC, Canada.
2
Genome Stability Laboratory, Oncology Division, CHU de Québec-Université Laval, Québec, QC, Canada.
3
Department of Molecular Biology, Medical Biochemistry and Pathology, Laval University Cancer Research Center, Québec, QC, Canada.

Abstract

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.

KEYWORDS:

Arginine methylation; DDX5; RGG/RG motif; RNA helicase; XRN2

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