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Topoisomerase IIα maintains genomic stability through decatenation G2 checkpoint signaling

Abstract

Topoisomerase IIα (topoIIα) is an essential mammalian enzyme that topologically modifies DNA and is required for chromosome segregation during mitosis. Previous research suggests that inhibition of topoII decatenatory activity triggers a G2 checkpoint response, which delays mitotic entry because of insufficient decatenation of daughter chromatids. Here we examine the effects of both topoIIα and topoIIβ on decatenatory activity in cell extracts, DNA damage and decatenation G2 checkpoint function, and the frequencies of p16INK4A allele loss and gain. In diploid human fibroblast lines, depletion of topoIIα by small-interfering RNA was associated with severely reduced decatenatory activity, delayed progression from G2 into mitosis and insensitivity to G2 arrest induced by the topoII catalytic inhibitor ICRF-193. Furthermore, interphase nuclei of topoIIα-depleted cells showed increased frequencies of losses and gains of the tumor suppressor genetic locus p16INK4A. This study shows that the topoIIα protein is required for decatenation G2 checkpoint function, and inactivation of decatenation and the decatenation G2 checkpoint leads to abnormal chromosome segregation and genomic instability.

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Abbreviations

topoII:

topoisomerase II

DSB:

double-strand break

NHDF:

normal human diploid fibroblast

NTC:

non-targeting control

MER:

mitotic entry rate

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Acknowledgements

Financial Support: This work was supported by an NIH-NCI Ruth L. Kirschstein National Research Service Award Grant # 1F32CA134155-01 (JJ Bower) and PHS grants CA81343 and P30-ES10126 (WK Kaufmann).

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Correspondence to W K Kaufmann.

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Bower, J., Karaca, G., Zhou, Y. et al. Topoisomerase IIα maintains genomic stability through decatenation G2 checkpoint signaling. Oncogene 29, 4787–4799 (2010). https://doi.org/10.1038/onc.2010.232

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Keywords

  • decatenation
  • G2 checkpoint
  • topoisomerase IIα
  • topoisomerase IIβ
  • chromosomal instability

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