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CIP2A mediates mitotic recruitment of SLX4/MUS81/XPF to replication stress-induced DNA lesions to maintain genome integrity

ABSTRACT: Perturbed DNA replication can result in genomic regions that remain incompletely replicated at the point that cells enter mitosis. Transmission of replication-mediated DNA lesions into mitosis may prevent proper chromosomes segregation, and cells therefore require mechanisms to resolve these lesions during mitosis. CIP2A in conjunction with TOPBP1 has been described to function as a molecular tether to connect broken DNA molecules during mitosis, but a role for CIP2A in processing of incompletely replicated DNA has remained unclear. We find that the CIP2A-TOPBP1 complex forms large filamentous structures at sites of at sites of mitotic DNA synthesis during mitosis. Using endogenous IP we aimed to identify CIP2A-interactions induced by irradiation or replication inhibition, and find that CIP2A recruits members the SMX tri-nuclease complex, including SLX4, MUS81 and ERCC1/XPF.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Epithelial Cell, Cell Culture

SUBMITTER: Rolf de Boer

LAB HEAD: Marcel van

PROVIDER: PXD059881 | Pride | 2025-10-23

REPOSITORIES: Pride

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			Action	DRS
	20230719_2023-77_CIP2A_KO_irradiation_A.raw	Raw
	20230719_2023-77_CIP2A_KO_irradiation_B.raw	Raw
	20230719_2023-77_CIP2A_KO_irradiation_C.raw	Raw
	20230719_2023-77_CIP2A_KO_untreated_A.raw	Raw
	20230719_2023-77_CIP2A_KO_untreated_B.raw	Raw

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Project description:Mitosis entails global alterations to chromosome structure and nuclear architecture, concomitant with transient silencing of transcription. How cells transmit transcriptional states through mitosis remains incompletely understood. While many nuclear factors dissociate from mitotic chromosomes, the observation that certain nuclear factors and chromatin features remain associated with individual loci during mitosis originated the hypothesis that they could provide transcriptional memory through mitosis. To obtain the first genome-wide view of the dynamics of chromatin structure during mitosis, we compared the DNase sensitivity of interphase and mitotic chromatin at two stages of cellular maturation in a rapidly dividingmurine erythroblastmodel. Despite global chromosome condensation visible during mitosis at the microscopic level, the chromatin accessibility landscape is largely unaltered. However, mitotic chromatin accessibility is locally dynamic, with individual loci maintaining none, some, or all of their interphase accessibility. Mitotic reduction in accessibility occurs primarily within narrow, highly hypersensitive sites that frequently coincide with transcription factor binding sites, whereas broader domains of moderate accessibility tend to be more stable. In mitosis, proximal promoters generally maintain their accessibility, whereas distal regulatory elements preferentially lose accessibility. Promoters with the highest degree of accessibility preservation in mitosis tend to also be accessible across many murine tissues in interphase. Transcription factor GATA1 exerts site-specific changes in interphase accessibility that are most pronounced at distal regulatory elements, but does not visibly influence mitotic accessibility. We conclude that features of open chromatin are remarkably stable through mitosis and are modulated at the level of individual genes and regulatory elements. Dnase-Seq data is integrated with Chip-seq [GSE36589, GSE30142] and RNA-seq to examine epigentic changes in mitosis. We performed DNase-seq on two cell lines, G1E and G1E-ER4, both on an asynchronus population, and on a sample of cells in mitosis; each of the 4 experiments in triplicate.

Dataset Information

CIP2A mediates mitotic recruitment of SLX4/MUS81/XPF to replication stress-induced DNA lesions to maintain genome integrity

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