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RFWD3-Dependent Ubiquitination of RPA Regulates Repair at Stalled Replication Forks
We have used quantitative proteomics to profile ubiquitination in the DNA damage response (DDR). We demonstrate that RPA, which functions as a protein scaffold in the replication stress response, is multiply ubiquitinated upon replication fork stalling. Ubiquitination of RPA occurs on chromatin, inv...
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Published in: | Molecular cell 2015-10, Vol.60 (2), p.280-293 |
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Main Authors: | , , , , , , , , , |
Format: | Article |
Language: | English |
Subjects: | |
Citations: | Items that this one cites Items that cite this one |
Online Access: | Get full text |
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Summary: | We have used quantitative proteomics to profile ubiquitination in the DNA damage response (DDR). We demonstrate that RPA, which functions as a protein scaffold in the replication stress response, is multiply ubiquitinated upon replication fork stalling. Ubiquitination of RPA occurs on chromatin, involves sites outside its DNA binding channel, does not cause proteasomal degradation, and increases under conditions of fork collapse, suggesting a role in repair at stalled forks. We demonstrate that the E3 ligase RFWD3 mediates RPA ubiquitination. RFWD3 is necessary for replication fork restart, normal repair kinetics during replication stress, and homologous recombination (HR) at stalled replication forks. Mutational analysis suggests that multisite ubiquitination of the entire RPA complex is responsible for repair at stalled forks. Multisite protein group sumoylation is known to promote HR in yeast. Our findings reveal a similar requirement for multisite protein group ubiquitination during HR at stalled forks in mammalian cells.
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•RPA is ubiquitinated on chromatin in response to replication fork stalling•Ubiquitination of RPA is mediated by the E3 ligase RFWD3•RFWD3 promotes replication fork restart and homologous recombination at stalled forks•Multisite ubiquitination of the entire RPA complex is necessary for HR
Using quantitative proteomics, Elia et al. demonstrate that RPA is multiply ubiquitinated in response to replication fork stalling. RPA ubiquitination is mediated by the E3 ligase RFWD3, which promotes homologous recombination at stalled forks and replication fork restart. |
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ISSN: | 1097-2765 1097-4164 |
DOI: | 10.1016/j.molcel.2015.09.011 |