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

iScience (Jul 2024)

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

Affiliations

Rebecca M. Jones: Institute of Cancer and Genomic Sciences, Birmingham Centre for Genome Biology, University of Birmingham, Birmingham, UK
Joaquin Herrero Ruiz: Institute of Cancer and Genomic Sciences, Birmingham Centre for Genome Biology, University of Birmingham, Birmingham, UK; Department of Genetics, The Graduate University for Advanced Studies (SOKENDAI), Mishima, Shizuoka, Japan
Shaun Scaramuzza: Institute of Cancer and Genomic Sciences, Birmingham Centre for Genome Biology, University of Birmingham, Birmingham, UK
Sarmi Nath: Institute of Cancer and Genomic Sciences, Birmingham Centre for Genome Biology, University of Birmingham, Birmingham, UK
Chaoyu Liu: Institute of Cancer and Genomic Sciences, Birmingham Centre for Genome Biology, University of Birmingham, Birmingham, UK
Marta Henklewska: Institute of Cancer and Genomic Sciences, Birmingham Centre for Genome Biology, University of Birmingham, Birmingham, UK
Toyoaki Natsume: Department of Chromosome Science, National Institute of Genetics, Research Organization of Information and Systems, Mishima, Shizuoka, Japan; Department of Genetics, The Graduate University for Advanced Studies (SOKENDAI), Mishima, Shizuoka, Japan
Robert G. Bristow: Cancer Research UK – Manchester Institute, Manchester Cancer Research Center, Manchester, UK
Francisco Romero: Department of Microbiology, University of Seville, Seville, Spain
Masato T. Kanemaki: Department of Chromosome Science, National Institute of Genetics, Research Organization of Information and Systems, Mishima, Shizuoka, Japan; Department of Genetics, The Graduate University for Advanced Studies (SOKENDAI), Mishima, Shizuoka, Japan; Department of Biological Science, The University of Tokyo, Tokyo, Japan
Agnieszka Gambus: Institute of Cancer and Genomic Sciences, Birmingham Centre for Genome Biology, University of Birmingham, Birmingham, UK; Corresponding author

Journal volume & issue: Vol. 27, no. 7
p. 110260

Abstract

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Summary: 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.

Published in iScience

ISSN: 2589-0042 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science
Website: http://www.cell.com/iscience/home

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