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The termination of UHRF1-dependent PAF15 ubiquitin signaling is regulated by USP7 and ATAD5

Ryota Miyashita1, Atsuya Nishiyama1, Weihua Qin2

  • 1Division of Cancer Cell Biology, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan.

Elife
|February 3, 2023
PubMed

Insights

Deubiquitylation by USP7 and ATAD5-mediated unloading terminate UHRF1-dependent PAF15 ubiquitin signaling. Completion of DNA methylation by DNMT1 is essential for removing PAF15Ub2 from chromatin.

Area of Science:

  • Epigenetics and DNA Methylation
  • Ubiquitin Signaling
  • Chromatin Biology

Background:

  • UHRF1-dependent ubiquitin signaling regulates maintenance DNA methylation during replication.
  • UHRF1 catalyzes PAF15 ubiquitylation (PAF15Ub2), controlling DNMT1 localization and activity.
  • Mechanisms for terminating this signaling and coordinating with DNA methylation completion remain unclear.

Purpose of the Study:

  • To elucidate the mechanisms terminating UHRF1-mediated PAF15 ubiquitin signaling.
  • To understand the coordination between signaling termination and DNA methylation completion.
  • To identify key factors involved in PAF15 removal from chromatin.

Main Methods:

  • Investigated protein interactions using co-immunoprecipitation.
  • Utilized depletion and inhibition strategies for USP7 and DNMT1.
  • Analyzed PAF15 localization and ubiquitylation status on chromatin via microscopy and Western blotting.
  • Assessed the role of ATAD5 in PAF15 removal.

Main Results:

  • USP7 specifically interacts with PAF15Ub2 in a DNMT1 complex on replicating chromatin.
  • USP7 depletion or disrupted USP7-PAF15 interaction causes PAF15Ub2 accumulation.
  • Non-ubiquitylated PAF15 (PAF15Ub0) is removed by ATAD5.
  • Inhibiting DNMT1 catalytic activity prevents PAF15Ub2 chromatin removal, indicating completion of DNA methylation is crucial.

Conclusions:

  • USP7-mediated deubiquitylation and ATAD5-mediated unloading are critical for PAF15 removal from chromatin.
  • The completion of maintenance DNA methylation by DNMT1 is essential for terminating UHRF1-mediated ubiquitin signaling.
  • This study provides molecular insights into the disassembly of the DNA methylation machinery at the end of S phase.
Keywords:
ATAD5DNA methylationDNMT1PAF15USP7cell biologyxenopus

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