Characterizing replisome disassembly in human cells

Rebecca M. Jones; Joaquin Herrero Ruiz; Shaun Scaramuzza; Sarmi Nath; Chaoyu Liu; Marta Henklewska; Toyoaki Natsume; Robert G. Bristow; Francisco Romero; Masato T. Kanemaki; Agnieszka Gambus

doi:10.1016/j.isci.2024.110260

Characterizing replisome disassembly in human cells

Rebecca M. Jones, Joaquin Herrero Ruiz, Shaun Scaramuzza, Sarmi Nath, Chaoyu Liu, Marta Henklewska, Toyoaki Natsume, Robert G. Bristow, Francisco Romero, Masato T. Kanemaki, Agnieszka Gambus^*

^*Corresponding author for this work

School of Biosciences

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

Abstract

To ensure timely duplication of the entire eukaryotic genome, thousands of replication machineries (replisomes) act on genomic DNA at any time during S phase. In the final stages of this process, replisomes are unloaded from chromatin. Unloading is driven by polyubiquitylation of MCM7, a subunit of the terminated replicative helicase, and processed by p97/VCP segregase. Most of our knowledge of replication termination comes from model organisms, and little is known about how this process is executed and regulated in human somatic cells. Here we show that replisome disassembly in this system requires CUL2^LRR1-driven MCM7 ubiquitylation, p97, and UBXN7 for unloading and provide evidence for “backup” mitotic replisome disassembly, demonstrating conservation of such mechanisms. Finally, we find that small-molecule inhibitors against Cullin ubiquitin ligases (CULi) and p97 (p97i) affect replisome unloading but also lead to induction of replication stress in cells, which limits their usefulness to specifically target replisome disassembly processes.

Original language	English
Article number	110260
Number of pages	26
Journal	iScience
Volume	27
Issue number	7
Early online date	12 Jun 2024
DOIs	https://doi.org/10.1016/j.isci.2024.110260
Publication status	Published - 19 Jul 2024

Bibliographical note

Data Access Statement

Further information and requests for resources and reagents should be directed to and will be fulfilled by the lead contact, Aga Gambus ([email protected]). Microscopy data reported in this paper will be shared by the lead contact upon request.

Keywords

biochemistry
cell biology
genetics

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10.1016/j.isci.2024.110260Licence: CC BY 4.0

RM Jones et al_Characterizing replisome disassembly in human cells
Copyright © 2024 The Author(s). Published by Elsevier Inc. This is an open access article under the CC BY license (https://creativecommons.org/licenses/by/4.0/).
Final published version, 5.74 MBLicence: CC BY 4.0

Cite this

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title = "Characterizing replisome disassembly in human cells",

abstract = "To ensure timely duplication of the entire eukaryotic genome, thousands of replication machineries (replisomes) act on genomic DNA at any time during S phase. In the final stages of this process, replisomes are unloaded from chromatin. Unloading is driven by polyubiquitylation of MCM7, a subunit of the terminated replicative helicase, and processed by p97/VCP segregase. Most of our knowledge of replication termination comes from model organisms, and little is known about how this process is executed and regulated in human somatic cells. Here we show that replisome disassembly in this system requires CUL2LRR1-driven MCM7 ubiquitylation, p97, and UBXN7 for unloading and provide evidence for “backup” mitotic replisome disassembly, demonstrating conservation of such mechanisms. Finally, we find that small-molecule inhibitors against Cullin ubiquitin ligases (CULi) and p97 (p97i) affect replisome unloading but also lead to induction of replication stress in cells, which limits their usefulness to specifically target replisome disassembly processes.",

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author = "Jones, {Rebecca M.} and Ruiz, {Joaquin Herrero} and Shaun Scaramuzza and Sarmi Nath and Chaoyu Liu and Marta Henklewska and Toyoaki Natsume and Bristow, {Robert G.} and Francisco Romero and Kanemaki, {Masato T.} and Agnieszka Gambus",

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AU - Jones, Rebecca M.

AU - Ruiz, Joaquin Herrero

AU - Scaramuzza, Shaun

AU - Nath, Sarmi

AU - Liu, Chaoyu

AU - Henklewska, Marta

AU - Natsume, Toyoaki

AU - Bristow, Robert G.

AU - Romero, Francisco

AU - Kanemaki, Masato T.

AU - Gambus, Agnieszka

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N2 - To ensure timely duplication of the entire eukaryotic genome, thousands of replication machineries (replisomes) act on genomic DNA at any time during S phase. In the final stages of this process, replisomes are unloaded from chromatin. Unloading is driven by polyubiquitylation of MCM7, a subunit of the terminated replicative helicase, and processed by p97/VCP segregase. Most of our knowledge of replication termination comes from model organisms, and little is known about how this process is executed and regulated in human somatic cells. Here we show that replisome disassembly in this system requires CUL2LRR1-driven MCM7 ubiquitylation, p97, and UBXN7 for unloading and provide evidence for “backup” mitotic replisome disassembly, demonstrating conservation of such mechanisms. Finally, we find that small-molecule inhibitors against Cullin ubiquitin ligases (CULi) and p97 (p97i) affect replisome unloading but also lead to induction of replication stress in cells, which limits their usefulness to specifically target replisome disassembly processes.

AB - To ensure timely duplication of the entire eukaryotic genome, thousands of replication machineries (replisomes) act on genomic DNA at any time during S phase. In the final stages of this process, replisomes are unloaded from chromatin. Unloading is driven by polyubiquitylation of MCM7, a subunit of the terminated replicative helicase, and processed by p97/VCP segregase. Most of our knowledge of replication termination comes from model organisms, and little is known about how this process is executed and regulated in human somatic cells. Here we show that replisome disassembly in this system requires CUL2LRR1-driven MCM7 ubiquitylation, p97, and UBXN7 for unloading and provide evidence for “backup” mitotic replisome disassembly, demonstrating conservation of such mechanisms. Finally, we find that small-molecule inhibitors against Cullin ubiquitin ligases (CULi) and p97 (p97i) affect replisome unloading but also lead to induction of replication stress in cells, which limits their usefulness to specifically target replisome disassembly processes.

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Characterizing replisome disassembly in human cells

Abstract

Bibliographical note

Data Access Statement

Keywords

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