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

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

  • bioRxiv. 2025 Jan 9:2025.01.09.632184. doi: 10.1101/2025.01.09.632184.
Lilly F Chiou 1 2,  Deepika Jayaprakash 2 3 4,  Gaith N Droby 1 2,  Xingyuan Zhang 2 5,  Yang Yang 2 6,  C Allie Mills 7,  Thomas S Webb 7,  Natalie K Barker 7,  Di Wu 8,  Laura E Herring 7,  Jessica Bowser 2 9,  Cyrus Vaziri 2 9 10
Affiliations

Affiliations

  • 1 Curriculum in Genetics and Molecular Biology, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA.
  • 2 Department of Pathology and Laboratory Medicine, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA.
  • 3 Oral and Craniofacial Biomedicine Program, Adams School of Dentistry, University of North Carolina at Chapel Hill, NC 27599, USA.
  • 4 Present address: Department of Biochemistry, Vanderbilt University School of Medicine, Nashville, TN 37237, USA.
  • 5 Present address: Department of Biostatistics and Bioinformatics, Duke University, Durham, NC, USA.
  • 6 Present address: In Vivo Neurobiology Group, Neurobiology Laboratory, National Institute of Environmental Health Sciences, National Institutes of Health, 111 T.W. Alexander Drive, Research Triangle Park, NC 27709, USA.
  • 7 UNC Metabolomics & Proteomics Core Facility, Department of Pharmacology, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA.
  • 8 Division of Oral and Craniofacial Health Science, Adams School of Dentistry, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA.
  • 9 UNC Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA.
  • 10 Lead Contact.
PMID: 39829812 DOI: 10.1101/2025.01.09.632184
Abstract

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-molecule DNA fiber analysis, we show that RNF25 protects reversed DNA replication forks generated by the fork remodeler HLTF from nucleolytic degradation by MRE11 and CtIP. Mechanistically, RNF25 interacts with the replication fork protection factor REV7 and recruits REV7 to nascent DNA after replication stress. The role of RNF25 in protecting replication forks is fully separable from its canonical functions in ubiquitin conjugation. This work reveals the RNF25-REV7 signaling axis as an important protective mechanism in cells experiencing replication stress.

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