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OTUD5 limits replication fork instability by organizing chromatin remodelers.

Angelo de Vivo1, Hongseon Song2, Yujin Lee2

  • 1Department of Molecular Biosciences, College of Arts and Sciences, University of South Florida, Tampa, FL 33647, USA.

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The deubiquitinase OTUD5 limits replication stress by organizing a complex that includes FACT and HDAC1/2. This interaction prevents excessive FACT loading and R-loop formation, maintaining genomic stability.

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Area of Science:

  • Molecular Biology
  • Genomics
  • Cell Biology

Background:

  • Replication fork progression is vital for genomic maintenance.
  • Transcription-replication conflicts can destabilize replication forks.
  • Chromatin remodelers modulate replication stress, but their organization is unclear.

Purpose of the Study:

  • To investigate the role of OTUD5 in regulating replication stress.
  • To identify the protein complex and mechanism by which OTUD5 limits replication stress.

Main Methods:

  • Recruitment assays to replication forks.
  • Analysis of replication fork stress markers.
  • Engineering of cell lines to uncouple protein interactions.
  • Proteomic analysis and pathway activation studies.

Main Results:

  • OTUD5 is recruited to replication forks and its depletion causes stress.
  • OTUD5 assembles a complex of FACT, HDAC1, and HDAC2 via its C-terminal tail.
  • Disrupting OTUD5-FACT interaction increases FACT loading, R-loop formation, and replication stress.
  • OTUD5 recruits HDAC1/2 to reduce H4K16 acetylation and FACT recruitment.
  • OTUD5 deficiency activates the Fanconi Anemia pathway for cell survival.

Conclusions:

  • OTUD5 plays a novel role in limiting transcription-induced replication stress.
  • A new interaction network involving OTUD5, FACT, HDAC1, and HDAC2 is identified.
  • This network is crucial for maintaining replication fork integrity.