Proteomics

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The E3 ubiquitin ligase RNF25 protects stalled DNA replication forks

ABSTRACT: Cancer cells experience high levels of replication stress and have therefore adapted mechanisms to tolerate replication stress. Using a CRISPR screen, we identified RNF25 as a mediator of replication stress tolerance in cancer cells. RNF25-ablation results in ssDNA accumulation, decreased replication fork movement, and increased replication fork degradation dependent on nucleases MRE11 and CtIP. To identify potential interactors of RNF25 in mediating replication stress tolerance, we performed mass-spectrometry analysis of immunopurified RNF25-complexes. We discovered REV7 as a novel binding partner of RNF25. REV7 was previously shown to protect replication forks in a Shieldin-independent but REV1- and REV3L-dependent manner. Further investigation revealed that RNF25 is also epistatic to REV1 and REV3L in a fork protection pathway and that REV7 recruitment to replication forks following stress is dependent on RNF25. Our findings establish a novel role of RNF25 in replication fork protection.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Lung

SUBMITTER: Lilly Chiou  

LAB HEAD: Cyrus Vaziri

PROVIDER: PXD056698 | Pride | 2025-06-30

REPOSITORIES: Pride

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Publications

The RING Finger E3 Ligase RNF25 Protects DNA Replication Forks Independently of its Canonical Roles in Ubiquitin Signaling.

Chiou Lilly F LF   Jayaprakash Deepika D   Droby Gaith N GN   Zhang Xingyuan X   Yang Yang Y   Mills C Allie CA   Webb Thomas S TS   Barker Natalie K NK   Wu Di D   Herring Laura E LE   Bowser Jessica J   Vaziri Cyrus C  

bioRxiv : the preprint server for biology 20250109

The DNA damage response (DDR) mechanisms that allow cells to tolerate DNA replication stress are critically important for genome stability and cell viability. Using an unbiased genetic screen we identify a role for the RING finger E3 ubiquitin ligase RNF25 in promoting DNA replication stress tolerance. In response to DNA replication stress, RNF25-deficient cells generate aberrantly high levels of single-stranded DNA (ssDNA), accumulate in S-phase and show reduced mitotic entry. Using single-mole  ...[more]

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