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PMID:22344029
| Citation |
Adamson, B, Smogorzewska, A, Sigoillot, FD, King, RW and Elledge, SJ (2012) A genome-wide homologous recombination screen identifies the RNA-binding protein RBMX as a component of the DNA-damage response. Nat. Cell Biol. 14:318-28 |
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| 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. |
| Links |
PubMed PMC3290715 Online version:10.1038/ncb2426 |
| Keywords |
BRCA2 Protein/genetics; BRCA2 Protein/metabolism; Cell Cycle Proteins/genetics; Cell Cycle Proteins/metabolism; Cell Line, Tumor; DNA Damage; DNA Repair; Gene Regulatory Networks; Genome, Human/genetics; Green Fluorescent Proteins/genetics; Green Fluorescent Proteins/metabolism; Heterogeneous-Nuclear Ribonucleoproteins/genetics; Heterogeneous-Nuclear Ribonucleoproteins/metabolism; Histone Chaperones/genetics; Histone Chaperones/metabolism; Homologous Recombination; Humans; Immunoblotting; Microscopy, Fluorescence; Models, Genetic; Nuclear Pore Complex Proteins/genetics; Nuclear Pore Complex Proteins/metabolism; Nuclear Proteins/genetics; Nuclear Proteins/metabolism; Poly(ADP-ribose) Polymerases/genetics; Poly(ADP-ribose) Polymerases/metabolism; RNA Interference; RNA Precursors/genetics; RNA Precursors/metabolism; RNA, Small Interfering/genetics; RNA-Binding Proteins/genetics; RNA-Binding Proteins/metabolism; Rad51 Recombinase/genetics; Rad51 Recombinase/metabolism; Reverse Transcriptase Polymerase Chain Reaction; Transcription Factors/genetics; Transcription Factors/metabolism |
| edit table |
Significance
Annotations
| Gene product | Qualifier | GO Term | Evidence Code | with/from | Aspect | Extension | Notes | Status |
|---|---|---|---|---|---|---|---|---|
| GO:0005654: nucleoplasm |
ECO:0000314: |
C |
Figure 3.b. and 3.c. indicate that four HIRIP3 siRNA causes HR defect. Even though these HIRIP3 siRNA are also correlated with off target Rad51 depletion, only siHIRIP3-5 had 7 nucleotide seed match to 3‘UTR Rad51 off-targeting. Therefore, HR defects are 3-fold enriched for antisense seed sequence complementary to Rad51. However, Figure 3.b. also indicates three siRNAs defect HR depleted Rad51 without full seed complementary to the 3’UTR. Therefore, HIRIP3 siRNAs can change the mechanisms of Rad51 off-targeting. |
complete | ||||
Notes
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References
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