The capacity of Topoisomerase II (Topo II) to remove DNA catenations that arise after replication is essential to ensure faithful chromosome segregation. Topo II activity is monitored during G2 by a specific checkpoint pathway that delays entry into mitosis until the chromosomes are properly decatenated. Recently, we demonstrated that the mitotic defects that are characteristic of cells depleted of MCPH1 function, a protein mutated in primary microcephaly, are not a consequence of a weakened G2 decatenation checkpoint response. However, the mitotic defects could be accounted for by a minor defect in the activity of Topo II during G2/M. To test this hypothesis, we have tracked at live single cell resolution the dynamics of mitosis in MCPH1 depleted HeLa cells upon catalytic inhibition of Topo II. Our analyses demonstrate that neither chromosome alignment nor segregation are more susceptible to minor perturbation in decatenation in MCPH1 deficient cells, as compared with control cells. Interestingly, MCPH1 depleted cells were more prone to mitotic cell death when decatenation was perturbed. Furthermore, when the G2 arrest that was induced by catalytic inhibition of Topo II was abrogated by Chk1 inhibition, the incidence of mitotic cell death was also increased. Taken together, our data suggest that the MCPH1 lack of function increases mitotic cell hypersensitivity to the catalytic inhibition of Topo II.
Bibliographical noteFunding Information:
Funding: This work was supported by Junta de ?ndalucía (Funding program ‘?yudas a grupos de investigación’, reference RNM-924) and Consejería de Salud, Junta de Andalucía (Grant number PI-0110-2017). Research in the laboratory of D. Clarke was financially supported by NIH grants R01GM112793 and R01GM130858.
Acknowledgments: The authors express their gratitude to Ryoko Kuriyama (University of Minnesota, USA) for helpful discussions, Nieves de la Casa (CICT, Universidad de Jaén, Spain) for technical assistance, and Cristina Cardoso (University of Darmstadt, Germany) for methodological advice on gH2AX analyses and general support. Technical and human support provided by CICT of Universidad de Jaén (UJA, MINECO, Junta de Andalucía, FEDER) is gratefully acknowledged.
- Anaphase errors
- Chromosome condensation
- Chromosome segregation
- Decatenation checkpoint
- Mitotic catastrophe
- Mitotic cell death
- Topoisomerase II
PubMed: MeSH publication types
- Journal Article
- Research Support, N.I.H., Extramural
- Research Support, Non-U.S. Gov't