Title | A genome-wide homologous recombination screen identifies the RNA-binding protein RBMX as a component of the DNA-damage response. |
Publication Type | Journal Article |
Year of Publication | 2012 |
Authors | Adamson, B, Smogorzewska, A, Sigoillot, FD, King, RW, Elledge, SJ |
Journal | Nat Cell Biol |
Volume | 14 |
Issue | 3 |
Pagination | 318-28 |
Date Published | 2012 Feb 19 |
ISSN | 1476-4679 |
Keywords | BRCA2 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 | <p>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.</p> |
DOI | 10.1038/ncb2426 |
Alternate Journal | Nat Cell Biol |
PubMed ID | 22344029 |
PubMed Central ID | PMC3290715 |
Grant List | T32CA09216 / CA / NCI NIH HHS / United States R37 GM044664-24 / GM / NIGMS NIH HHS / United States T32 CA009216 / CA / NCI NIH HHS / United States R37 GM044664 / GM / NIGMS NIH HHS / United States / HHMI / Howard Hughes Medical Institute / United States R01 GM044664 / GM / NIGMS NIH HHS / United States |