Abstract
DNA double-strand breaks (DSBs) are repaired mainly by non-homologous end joining or homologous recombination (HR). Cell cycle stage and DNA end resection are believed to regulate the commitment to HR repair. Here we identify RNF138 as a ubiquitin E3 ligase that regulates the HR pathway. RNF138 is recruited to DNA damage sites through zinc fingers that have a strong preference for DNA with 5'- or 3'-single-stranded overhangs. RNF138 stimulates DNA end resection and promotes ATR-dependent signalling and DSB repair by HR, thereby contributing to cell survival on exposure to DSB-inducing agents. Finally, we establish that RNF138-dependent Ku removal from DNA breaks is one mechanism whereby RNF138 can promote HR. These results establish RNF138 as an important regulator of DSB repair pathway choice.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Ataxia Telangiectasia Mutated Proteins / metabolism
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Cell Line, Tumor
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DNA Breaks, Double-Stranded*
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DNA End-Joining Repair
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DNA Helicases / genetics
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DNA Helicases / metabolism*
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DNA Repair*
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DNA, Neoplasm / genetics
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DNA, Neoplasm / metabolism*
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DNA, Single-Stranded / genetics
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DNA, Single-Stranded / metabolism
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HEK293 Cells
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HeLa Cells
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Humans
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Immunoblotting
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Ku Autoantigen
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Luminescent Proteins / genetics
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Luminescent Proteins / metabolism
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MCF-7 Cells
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Microscopy, Confocal
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Mutation
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Protein Binding
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RNA Interference
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Recombinational DNA Repair
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Ubiquitin-Protein Ligases / genetics
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Ubiquitin-Protein Ligases / metabolism*
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Ubiquitination
Substances
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DNA, Neoplasm
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DNA, Single-Stranded
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Luminescent Proteins
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RNF138 protein, human
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Ubiquitin-Protein Ligases
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ATR protein, human
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Ataxia Telangiectasia Mutated Proteins
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DNA Helicases
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XRCC5 protein, human
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Ku Autoantigen