USP37 prevents premature disassembly of stressed replisomes by TRAIP

Abstract The eukaryotic replisome, which consists of the CDC45-MCM2-7-GINS (CMG) helicase, replicative polymerases, and several accessory factors, sometimes encounters proteinaceous obstacles that threaten genome integrity. These obstacles are targeted for removal or proteolysis by the E3 ubiquitin...

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Main Authors: Olga V. Kochenova, Giuseppina D’Alessandro, Domenic Pilger, Ernst Schmid, Sean L. Richards, Marcos Rios Garcia, Satpal S. Jhujh, Andrea Voigt, Vipul Gupta, Christopher J. Carnie, R. Alex Wu, Nadia Gueorguieva, Simon Lam, Grant S. Stewart, Johannes C. Walter, Stephen P. Jackson
Format: Article
Language:English
Published: Nature Portfolio 2025-06-01
Series:Nature Communications
Online Access:https://doi.org/10.1038/s41467-025-60139-z
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author Olga V. Kochenova
Giuseppina D’Alessandro
Domenic Pilger
Ernst Schmid
Sean L. Richards
Marcos Rios Garcia
Satpal S. Jhujh
Andrea Voigt
Vipul Gupta
Christopher J. Carnie
R. Alex Wu
Nadia Gueorguieva
Simon Lam
Grant S. Stewart
Johannes C. Walter
Stephen P. Jackson
author_facet Olga V. Kochenova
Giuseppina D’Alessandro
Domenic Pilger
Ernst Schmid
Sean L. Richards
Marcos Rios Garcia
Satpal S. Jhujh
Andrea Voigt
Vipul Gupta
Christopher J. Carnie
R. Alex Wu
Nadia Gueorguieva
Simon Lam
Grant S. Stewart
Johannes C. Walter
Stephen P. Jackson
author_sort Olga V. Kochenova
collection DOAJ
description Abstract The eukaryotic replisome, which consists of the CDC45-MCM2-7-GINS (CMG) helicase, replicative polymerases, and several accessory factors, sometimes encounters proteinaceous obstacles that threaten genome integrity. These obstacles are targeted for removal or proteolysis by the E3 ubiquitin ligase TRAIP, which associates with the replisome. However, TRAIP must be carefully regulated to avoid inappropriate ubiquitylation and disassembly of the replisome. Here, we demonstrate that human cells lacking the de-ubiquitylating enzyme USP37 are hypersensitive to topoisomerase poisons and other replication stress-inducing agents. Furthermore, TRAIP loss rescues the hypersensitivity of USP37 knockout cells to topoisomerase inhibitors. In Xenopus egg extracts depleted of USP37, TRAIP promotes premature CMG ubiquitylation and disassembly when converging replisomes stall. Finally, guided by AlphaFold-Multimer, we discovered that binding to CDC45 mediates USP37’s response to topological stress. We propose that USP37 protects genome stability by preventing TRAIP-dependent CMG unloading when replication stress impedes timely termination.
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spelling doaj-art-0909bef6c62a4638bfca3cba93ff67212025-08-20T02:37:33ZengNature PortfolioNature Communications2041-17232025-06-0116111710.1038/s41467-025-60139-zUSP37 prevents premature disassembly of stressed replisomes by TRAIPOlga V. Kochenova0Giuseppina D’Alessandro1Domenic Pilger2Ernst Schmid3Sean L. Richards4Marcos Rios Garcia5Satpal S. Jhujh6Andrea Voigt7Vipul Gupta8Christopher J. Carnie9R. Alex Wu10Nadia Gueorguieva11Simon Lam12Grant S. Stewart13Johannes C. Walter14Stephen P. Jackson15Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Blavatnik InstituteCancer Research UK Cambridge Institute, Li Ka Shing Building, Robinson WayThe Gurdon Institute and Department of Biochemistry, University of CambridgeDepartment of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Blavatnik InstituteCancer Research UK Cambridge Institute, Li Ka Shing Building, Robinson WayInstitute of Cancer and Genomic Sciences, College of Medical and Dental Sciences, University of BirminghamInstitute of Cancer and Genomic Sciences, College of Medical and Dental Sciences, University of BirminghamCancer Research UK Cambridge Institute, Li Ka Shing Building, Robinson WayCancer Research UK Cambridge Institute, Li Ka Shing Building, Robinson WayCancer Research UK Cambridge Institute, Li Ka Shing Building, Robinson WayDepartment of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Blavatnik InstituteCancer Research UK Cambridge Institute, Li Ka Shing Building, Robinson WayCancer Research UK Cambridge Institute, Li Ka Shing Building, Robinson WayInstitute of Cancer and Genomic Sciences, College of Medical and Dental Sciences, University of BirminghamDepartment of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Blavatnik InstituteCancer Research UK Cambridge Institute, Li Ka Shing Building, Robinson WayAbstract The eukaryotic replisome, which consists of the CDC45-MCM2-7-GINS (CMG) helicase, replicative polymerases, and several accessory factors, sometimes encounters proteinaceous obstacles that threaten genome integrity. These obstacles are targeted for removal or proteolysis by the E3 ubiquitin ligase TRAIP, which associates with the replisome. However, TRAIP must be carefully regulated to avoid inappropriate ubiquitylation and disassembly of the replisome. Here, we demonstrate that human cells lacking the de-ubiquitylating enzyme USP37 are hypersensitive to topoisomerase poisons and other replication stress-inducing agents. Furthermore, TRAIP loss rescues the hypersensitivity of USP37 knockout cells to topoisomerase inhibitors. In Xenopus egg extracts depleted of USP37, TRAIP promotes premature CMG ubiquitylation and disassembly when converging replisomes stall. Finally, guided by AlphaFold-Multimer, we discovered that binding to CDC45 mediates USP37’s response to topological stress. We propose that USP37 protects genome stability by preventing TRAIP-dependent CMG unloading when replication stress impedes timely termination.https://doi.org/10.1038/s41467-025-60139-z
spellingShingle Olga V. Kochenova
Giuseppina D’Alessandro
Domenic Pilger
Ernst Schmid
Sean L. Richards
Marcos Rios Garcia
Satpal S. Jhujh
Andrea Voigt
Vipul Gupta
Christopher J. Carnie
R. Alex Wu
Nadia Gueorguieva
Simon Lam
Grant S. Stewart
Johannes C. Walter
Stephen P. Jackson
USP37 prevents premature disassembly of stressed replisomes by TRAIP
Nature Communications
title USP37 prevents premature disassembly of stressed replisomes by TRAIP
title_full USP37 prevents premature disassembly of stressed replisomes by TRAIP
title_fullStr USP37 prevents premature disassembly of stressed replisomes by TRAIP
title_full_unstemmed USP37 prevents premature disassembly of stressed replisomes by TRAIP
title_short USP37 prevents premature disassembly of stressed replisomes by TRAIP
title_sort usp37 prevents premature disassembly of stressed replisomes by traip
url https://doi.org/10.1038/s41467-025-60139-z
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