USP37 protects mammalian cells during DNA replication stress by counteracting CUL2LRR1 and TRAIP

Fabrizio Villa; Johanna Ainsworth; Karim Labib

doi:10.1016/j.celrep.2025.115739

USP37 protects mammalian cells during DNA replication stress by counteracting CUL2^LRR1 and TRAIP

Fabrizio Villa
, Johanna Ainsworth
, Karim Labib (Lead / Corresponding author)

Genome Integrity

Research output: Contribution to journal › Article › peer-review

3 Citations (Scopus)

63 Downloads (Pure)

Abstract

The USP37 deubiquitylase is essential for mammalian cells to survive DNA replication stress, but the underlying mechanisms are unknown. Here, we demonstrate that USP37 binds the CDC45-MCM-GINS (CMG) helicase, which forms the stable core of the replisome until DNA replication termination when CMG is ubiquitylated and disassembled. USP37 contacts CDC45, and structure-guided mutations that displace USP37 from CMG cause sensitivity to DNA synthesis defects or topological stress. Binding to CDC45 at replication forks enables USP37 to counteract CMG ubiquitylation by the CUL2 ^LRR1 ligase, which subsequently induces replisome disassembly during termination. Correspondingly, depletion of CUL2 ^LRR1 suppresses the sensitivity of Usp37 mutants to DNA synthesis defects and ATR checkpoint kinase inhibitors. In contrast, mutation of the TRAIP ubiquitin ligase specifically suppresses the sensitivity of Usp37 mutants to topological stress. We propose that USP37 protects mammalian cells from replication stress by reversing the untimely action of the CUL2 ^LRR1 and TRAIP ubiquitin ligases.

Original language	English
Article number	115739
Number of pages	23
Journal	Cell Reports
Volume	44
Issue number	6
Early online date	22 May 2025
DOIs	https://doi.org/10.1016/j.celrep.2025.115739
Publication status	Published - 24 Jun 2025

Keywords

USP37
CUL2LRR1
TRAIP
DNA replication
CMG helicase
Ubiquitylation
Deubiquitylase
ATR checkpoint kinase
CUL2
CP: Molecular biology
deubiquitylase
ubiquitylation

ASJC Scopus subject areas

General Biochemistry,Genetics and Molecular Biology

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10.1016/j.celrep.2025.115739Licence: CC BY

Final Published VersionFinal published version, 27.1 MBLicence: CC BY

USP37 protects mammalian cells during DNA replication stress by counteracting CUL2LRR1 and TRAIP
Villa, F., Ainsworth, J. & Labib, K. P. M. (Lead / Corresponding author), 3 Sept 2024, BioRxiv, 80 p.
Research output: Working paper/Preprint › Preprint

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title = "USP37 protects mammalian cells during DNA replication stress by counteracting CUL2LRR1 and TRAIP",

abstract = "The USP37 deubiquitylase is essential for mammalian cells to survive DNA replication stress, but the underlying mechanisms are unknown. Here, we demonstrate that USP37 binds the CDC45-MCM-GINS (CMG) helicase, which forms the stable core of the replisome until DNA replication termination when CMG is ubiquitylated and disassembled. USP37 contacts CDC45, and structure-guided mutations that displace USP37 from CMG cause sensitivity to DNA synthesis defects or topological stress. Binding to CDC45 at replication forks enables USP37 to counteract CMG ubiquitylation by the CUL2 LRR1 ligase, which subsequently induces replisome disassembly during termination. Correspondingly, depletion of CUL2 LRR1 suppresses the sensitivity of Usp37 mutants to DNA synthesis defects and ATR checkpoint kinase inhibitors. In contrast, mutation of the TRAIP ubiquitin ligase specifically suppresses the sensitivity of Usp37 mutants to topological stress. We propose that USP37 protects mammalian cells from replication stress by reversing the untimely action of the CUL2 LRR1 and TRAIP ubiquitin ligases.",

keywords = "USP37, CUL2LRR1, TRAIP, DNA replication, CMG helicase, Ubiquitylation, Deubiquitylase, ATR checkpoint kinase, CUL2, CP: Molecular biology, deubiquitylase, ubiquitylation",

author = "Fabrizio Villa and Johanna Ainsworth and Karim Labib",

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AU - Villa, Fabrizio

AU - Ainsworth, Johanna

AU - Labib, Karim

PY - 2025/6/24

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N2 - The USP37 deubiquitylase is essential for mammalian cells to survive DNA replication stress, but the underlying mechanisms are unknown. Here, we demonstrate that USP37 binds the CDC45-MCM-GINS (CMG) helicase, which forms the stable core of the replisome until DNA replication termination when CMG is ubiquitylated and disassembled. USP37 contacts CDC45, and structure-guided mutations that displace USP37 from CMG cause sensitivity to DNA synthesis defects or topological stress. Binding to CDC45 at replication forks enables USP37 to counteract CMG ubiquitylation by the CUL2 LRR1 ligase, which subsequently induces replisome disassembly during termination. Correspondingly, depletion of CUL2 LRR1 suppresses the sensitivity of Usp37 mutants to DNA synthesis defects and ATR checkpoint kinase inhibitors. In contrast, mutation of the TRAIP ubiquitin ligase specifically suppresses the sensitivity of Usp37 mutants to topological stress. We propose that USP37 protects mammalian cells from replication stress by reversing the untimely action of the CUL2 LRR1 and TRAIP ubiquitin ligases.

AB - The USP37 deubiquitylase is essential for mammalian cells to survive DNA replication stress, but the underlying mechanisms are unknown. Here, we demonstrate that USP37 binds the CDC45-MCM-GINS (CMG) helicase, which forms the stable core of the replisome until DNA replication termination when CMG is ubiquitylated and disassembled. USP37 contacts CDC45, and structure-guided mutations that displace USP37 from CMG cause sensitivity to DNA synthesis defects or topological stress. Binding to CDC45 at replication forks enables USP37 to counteract CMG ubiquitylation by the CUL2 LRR1 ligase, which subsequently induces replisome disassembly during termination. Correspondingly, depletion of CUL2 LRR1 suppresses the sensitivity of Usp37 mutants to DNA synthesis defects and ATR checkpoint kinase inhibitors. In contrast, mutation of the TRAIP ubiquitin ligase specifically suppresses the sensitivity of Usp37 mutants to topological stress. We propose that USP37 protects mammalian cells from replication stress by reversing the untimely action of the CUL2 LRR1 and TRAIP ubiquitin ligases.

KW - USP37

KW - CUL2LRR1

KW - TRAIP

KW - DNA replication

KW - CMG helicase

KW - Ubiquitylation

KW - Deubiquitylase

KW - ATR checkpoint kinase

KW - CUL2

KW - CP: Molecular biology

KW - deubiquitylase

KW - ubiquitylation

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DO - 10.1016/j.celrep.2025.115739

M3 - Article

C2 - 40411782

SN - 2211-1247

VL - 44

JO - Cell Reports

JF - Cell Reports

IS - 6

M1 - 115739

ER -

USP37 protects mammalian cells during DNA replication stress by counteracting CUL2LRR1 and TRAIP

Abstract

Keywords

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