Promotion of RAD51-Mediated Homologous DNA Pairing by the RAD51AP1-UAF1 Complex

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Fengshan Liang1, Simonne Longerich1, Adam S Miller1

  • 1Department of Molecular Biophysics and Biochemistry, Yale University School of Medicine, New Haven, CT 06520, USA.

Cell Reports
|May 31, 2016
PubMed

Insights

UAF1 binds DNA and enhances DNA repair by homologous recombination through a complex with RAD51AP1 and RAD51. This reveals a novel, USP1-independent role for UAF1 in maintaining genome stability.

Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • The Fanconi anemia pathway is crucial for DNA damage response and homologous recombination repair.
  • UAF1, as part of the UAF1-USP1 complex, deubiquitinates FANCD2, impacting DNA repair.
  • UAF1 depletion leads to persistent FANCD2 ubiquitination, DNA damage hypersensitivity, and impaired homologous recombination.

Purpose of the Study:

  • To investigate the role of UAF1 in DNA repair beyond its known interaction with USP1.
  • To elucidate the mechanism by which UAF1 participates in homologous recombination.
  • To identify novel protein interactions and functions of UAF1 in genome maintenance.

Main Methods:

  • Biochemical assays to study protein-DNA interactions.
  • Co-immunoprecipitation to analyze protein complex formation.
  • In vitro assays to assess RAD51-mediated homologous DNA pairing.
  • Cellular studies to determine the biological significance of protein complexes.

Main Results:

  • UAF1 binds DNA and forms distinct complexes with RAD51AP1 and RAD51 via RAD51AP1.
  • Complex formation is mediated by SUMO-like domains in UAF1 and an SiM in RAD51AP1.
  • UAF1 enhances RAD51-mediated homologous DNA pairing, dependent on RAD51AP1 but independent of USP1.
  • The RAD51AP1-UAF1 complex facilitates the assembly of the RAD51 synaptic complex.

Conclusions:

  • UAF1 plays a critical role in homologous recombination by promoting synaptic complex formation.
  • This function of UAF1 is independent of its deubiquitinating activity via USP1.
  • The findings reveal a novel, USP1-independent mechanism for UAF1 in genome maintenance and DNA repair.

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