A genome-wide homologous recombination screen identifies the RNA-binding protein RBMX as a component of the DNA-damage response.

TitleA genome-wide homologous recombination screen identifies the RNA-binding protein RBMX as a component of the DNA-damage response.
Publication TypeJournal Article
Year of Publication2012
AuthorsAdamson, B, Smogorzewska, A, Sigoillot, FD, King, RW, Elledge, SJ
JournalNat Cell Biol
Volume14
Issue3
Pagination318-28
Date Published2012 Feb 19
ISSN1476-4679
KeywordsBRCA2 Protein, Cell Cycle Proteins, Cell Line, Tumor, DNA Damage, DNA Repair, Gene Regulatory Networks, Genome, Human, Green Fluorescent Proteins, Heterogeneous-Nuclear Ribonucleoproteins, Histone Chaperones, Homologous Recombination, Humans, Immunoblotting, Microscopy, Fluorescence, Models, Genetic, Nuclear Pore Complex Proteins, Nuclear Proteins, Poly(ADP-ribose) Polymerases, Rad51 Recombinase, Reverse Transcriptase Polymerase Chain Reaction, RNA Interference, RNA Precursors, RNA, Small Interfering, RNA-Binding Proteins, Transcription Factors
Abstract

Repair of DNA double-strand breaks is critical to genomic stability and the prevention of developmental disorders and cancer. A central pathway for this repair is homologous recombination (HR). Most knowledge of HR is derived from work in prokaryotic and eukaryotic model organisms. We carried out a genome-wide siRNA-based screen in human cells. Among positive regulators of HR we identified networks of DNA-damage-response and pre-mRNA-processing proteins, and among negative regulators we identified a phosphatase network. Three candidate proteins localized to DNA lesions, including RBMX, a heterogeneous nuclear ribonucleoprotein that has a role in alternative splicing. RBMX accumulated at DNA lesions through multiple domains in a poly(ADP-ribose) polymerase 1-dependent manner and promoted HR by facilitating proper BRCA2 expression. Our screen also revealed that off-target depletion of RAD51 is a common source of RNAi false positives, raising a cautionary note for siRNA screens and RNAi-based studies of HR.

DOI10.1038/ncb2426
Alternate JournalNat. Cell Biol.
PubMed ID22344029
PubMed Central IDPMC3290715
Grant ListT32CA09216 / CA / NCI NIH HHS / United States
R01 GM044664 / GM / NIGMS NIH HHS / United States
R37 GM044664-24 / GM / NIGMS NIH HHS / United States
T32 CA009216 / CA / NCI NIH HHS / United States
/ / Howard Hughes Medical Institute / United States
R37 GM044664 / GM / NIGMS NIH HHS / United States