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Related Experiment Video

Updated: Jan 18, 2026

Real-time Observation of the DNA Strand Exchange Reaction Mediated by Rad51
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UHRF1 Controls the Timing of RAD51 Removal During DNA Damage Repair Through Suppressing RFWD3.

Guangxue Liu1, Kaiyan Huang2, Shiyao Liu2

  • 1Zhejiang Provincial Key Laboratory of Pancreatic Disease, The First Affiliated Hospital of Zhejiang University, Hangzhou, Zhejiang, 310003, China.

Advanced Science (Weinheim, Baden-Wurttemberg, Germany)
|September 12, 2025
PubMed
Summary

UHRF1 protects RAD51 recombinase from degradation by RFWD3 E3 ligase. This interaction forms a negative feedback loop, ensuring RAD51 levels are maintained for DNA repair.

Keywords:
DNA damageRAD51RFWD3UHRF1

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Area of Science:

  • Molecular Biology
  • DNA Repair Mechanisms
  • Protein Regulation

Background:

  • RAD51 is crucial for homologous recombination (HR) repair of DNA double-strand breaks.
  • RFWD3 (an E3 ubiquitin ligase) removes RAD51 post-HR completion via ubiquitination and degradation.
  • The mechanism preventing premature RFWD3 activity on RAD51 was previously unknown.

Purpose of the Study:

  • To elucidate the regulatory mechanism protecting RAD51 from RFWD3.
  • To identify factors controlling RAD51 levels during DNA repair.
  • To characterize the feedback circuit involving RAD51, RFWD3, and UHRF1.

Main Methods:

  • Investigated the interaction between UHRF1, RAD51, and RFWD3.
  • Utilized ubiquitination assays to assess RFWD3 modification.
  • Examined the role of UHRF1 phosphorylation and PP4 phosphatase activity.

Main Results:

  • UHRF1 acts as an E3 ubiquitin ligase for RFWD3, thereby protecting RAD51.
  • RAD51 reciprocally protects RFWD3 from UHRF1, establishing a negative feedback loop.
  • UHRF1 phosphorylation status and PP4 activity regulate RFWD3 ubiquitination.

Conclusions:

  • A novel regulatory circuit involving UHRF1, RFWD3, and RAD51 maintains RAD51 levels for effective HR repair.
  • This feedback mechanism prevents premature RAD51 degradation, ensuring its availability for DNA repair.
  • Phosphorylation and dephosphorylation events fine-tune RFWD3 ubiquitination and RAD51 availability.