Single cell tracking reveals that msh2 is a key component of an early-acting dna damage-activated g2 checkpoint

ABSTRACT Dysfunction of cell-cycle checkpoints in DNA mismatch repair (MMR)-deficient cells in response to DNA damage has implications for anticancer therapy and genetic instability. We have

studied the cell-cycle effects of MMR deficiency (Msh2−/−) in primary mouse embryonic fibroblasts (MEFs) exposed to cisplatin (10 _μ_ M × 1 h) using time-lapse microscopy. Kinetic responses

of MEFs from different embryos and passage ages varied, but we report a consistent drug-induced inhibition of mitotic entry (approx. 50%). There was a loss of an early-acting (<5 h)

delay in G2 to M transition in Msh2−/− cells, although a later-acting G2 arrest was apparently normal. This suggests that Msh2 primarily acts to delay mitotic entry of cells already in G2,

that is, DNA damage incurred during G2 does not influence the cell once committed to mitotic traverse. Irrespective of Msh2 status, cisplatin treatment and the incurred DNA damage did not

effect mitotic traverse or show any evidence for early (within 24 h) cell death. The results indicate that Msh2−/− status can result in the premature commitment to mitosis of a cell

subpopulation, determined by the fraction residing in G2 at the time of damage induction. The findings suggest a new route to MMR-driven genetic instability that does not rely primarily on

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FUNCTIONS AS A TIMER REPORTING THE LENGTH OF MITOSIS Article Open access 09 January 2025 EMBRYONIC GENOME INSTABILITY UPON DNA REPLICATION TIMING PROGRAM EMERGENCE Article Open access 28

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and Kinetic Imaging Ltd. AUTHOR INFORMATION AUTHORS AND AFFILIATIONS * Department of Pathology, University of Wales College of Medicine, Heath Park, Cardiff, CF14 4XN, UK Nuria Marquez, 

Sally C Chappell & Paul J Smith * Cardiff School of Bioscience, Cardiff University, Cardiff, CF10 3US, UK Owen J Sansom & Alan R Clarke * Cancer Research Wales Laboratory, Velindre

NHS Trust, Cardiff, CF14 2TL, UK Jon Court * Department of Medical Biochemistry, University of Wales College of Medicine, Cardiff, UK Rachel J Errington Authors * Nuria Marquez View author

publications You can also search for this author inPubMed Google Scholar * Sally C Chappell View author publications You can also search for this author inPubMed Google Scholar * Owen J

Sansom View author publications You can also search for this author inPubMed Google Scholar * Alan R Clarke View author publications You can also search for this author inPubMed Google

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PERMISSIONS Reprints and permissions ABOUT THIS ARTICLE CITE THIS ARTICLE Marquez, N., Chappell, S., Sansom, O. _et al._ Single cell tracking reveals that Msh2 is a key component of an

early-acting DNA damage-activated G2 checkpoint. _Oncogene_ 22, 7642–7648 (2003). https://doi.org/10.1038/sj.onc.1206876 Download citation * Received: 20 March 2003 * Revised: 03 June 2003 *

Accepted: 04 June 2003 * Published: 23 October 2003 * Issue Date: 23 October 2003 * DOI: https://doi.org/10.1038/sj.onc.1206876 SHARE THIS ARTICLE Anyone you share the following link with

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content-sharing initiative KEYWORDS * G2 checkpoint * Msh2 * mouse embryonic fibroblasts * time-lapse microscopy * event analysis