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PCNA-binding activity separates RNF168 functions in DNA replication and DNA double-stranded break signaling.
Yang Yang1, Deepika Jayaprakash1,2, Satpal S Jhujh3
1Department of Pathology and Laboratory Medicine, University of North Carolina at Chapel Hill, 614 Brinkhous-Bullitt Building, 160 Medical Drive, Chapel Hill, NC 27599, USA.
Nucleic Acids Research
|October 24, 2024
Summary
RNF168 protein plays a dual role in DNA repair and replication. Researchers found a new mechanism where RNF168 supports DNA replication independently of its DNA damage response functions.
Area of Science:
- Molecular Biology
- DNA Replication
- DNA Damage Response
Background:
- RNF168 is known for its role in DNA double-strand break (DSB) repair by recruiting factors like 53BP1.
- Its involvement in DNA replication fork progression is suggested but not fully understood.
Purpose of the Study:
- To elucidate the precise role of RNF168 in DNA replication.
- To investigate the mechanism of RNF168 recruitment to replication factories.
- To determine if RNF168's functions in replication and DSB signaling are linked.
Main Methods:
- Investigated RNF168 recruitment to replication factories independently of ATM/RNF8.
- Identified a degenerate PCNA-interacting peptide (DPIP) motif and MIU domain in RNF168.
- Utilized an RNF168 mutant (ΔDPIP/ΔMIU1) deficient in PCNA interaction.
- Assessed the mutant's ability to support DNA replication and rescue 53BP1 foci formation in RNF168-/- cells.
Main Results:
- RNF168 is recruited to replication factories via interaction with mono-ubiquitylated PCNA through its DPIP motif and MIU domain.
- RNF168 recruitment to replication factories is independent of the canonical DSB response pathway.
- The RNF168 ΔDPIP/ΔMIU1 mutant fails to support DNA replication but fully rescues DSB-induced 53BP1 foci formation.
- RNF168's functions in DNA replication and DSB signaling are separable.
Conclusions:
- RNF168 promotes DNA replication through a novel mechanism involving PCNA interaction.
- This replication function is distinct from its established role in DNA double-strand break signaling.
- The study defines separable functions for RNF168 in maintaining genome stability during replication and repair.
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