Cited by 433DNA Repair MechanismsPARP inhibition in cancer therapyRNA Research and Splicing

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

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Adamson, Britt, Smogorzewska, Agata, Sigoillot, Frederic D., King, Randall W., Elledge, Stephen J.·2012·Nature Cell Biology

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.

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  • CRISPRi avoids the widespread off-target effects that are intrinsic to RNAi-based screening.

    “CRISPRi represents an important advance over RNAi, since it lacks the pervasive off-target effects (Gilbert et al., 2014) inherent to RNAi-based screening approaches (Adamson et al., 2012; Jackson et al., 2003; Kaelin, 2012).”

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Authors
Adamson, B., Smogorzewska, A., Sigoillot, F. D., King, R. W., & Elledge, S. J.
Affiliations
  • Brigham and Women's Hospital (US)
  • Harvard University (US)
Corresponding author
Elledge, Stephen J.
Venue
Nature Cell Biology
Published
2012-02-19
Volume
14
Issue
3
Pages
318-328
ISSN
  • 1465-7392 (ISSN-L)
  • 1476-4679
OpenAlex
W2052973047
Cited by
433
References
36
Citations by year
2026: 10 · 2025: 19 · 2024: 25 · 2023: 34 · 2022: 28 · 2021: 43 · 2020: 40 · 2019: 33 · 2018: 30 · 2017: 30 · 2016: 30 · 2015: 33 · 2014: 34 · 2013: 35 · 2012: 8
FWCI
14.15
Type
journal article
Language
en
Open access
Locations
  • Nature Cell Biology (publishedVersion)
  • PubMed (publishedVersion)
  • Europe PMC (PubMed Central) (acceptedVersion)
  • PubMed Central (acceptedVersion)
Funders
  • Howard Hughes Medical Institute
  • National Cancer Institute (T32CA09216, T32CA009216)
  • National Institute of General Medical Sciences (R37GM044664, R01GM044664)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA Repair Mechanisms
  • Health Sciences › Medicine › Oncology › PARP inhibition in cancer therapy
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Research and Splicing
Keywords
homologous recombination, RBMX, DNA damage response, DNA double-strand break repair, siRNA screen, off-target effects, RAD51, PARP1, BRCA2, pre-mRNA processing, RNA-binding proteins, alternative splicing, heterogeneous nuclear ribonucleoproteins
MeSH terms
DNA Damage, DNA Repair, Humans, Microscopy, Fluorescence, Models, Genetic, Nuclear Proteins, Poly(ADP-ribose) Polymerases, RNA Precursors, Transcription Factors, Immunoblotting, Genome, Human, RNA-Binding Proteins, Cell Cycle Proteins, Reverse Transcriptase Polymerase Chain Reaction, BRCA2 Protein, Nuclear Pore Complex Proteins, Heterogeneous-Nuclear Ribonucleoproteins, RNA Interference, RNA, Small Interfering, Cell Line, Tumor, Green Fluorescent Proteins, Rad51 Recombinase, Gene Regulatory Networks, Histone Chaperones, Homologous Recombination

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