Research Paper Volume 11, Issue 23 pp 11686—11721

Cytological and genetic consequences for the progeny of a mitotic catastrophe provoked by Topoisomerase II deficiency

Cristina Ramos-Pérez 1, 2, 6, , Margaret Dominska 3, , Laura Anaissi-Afonso 1, 2, , Sara Cazorla-Rivero 1, 2, , Oliver Quevedo 1, 7, , Isabel Lorenzo-Castrillejo 1, , Thomas D. Petes 3, , Félix Machín 1, 4, 5, ,

  • 1 Unidad de Investigación, Hospital Universitario Nuestra Señora de Candelaria, Santa Cruz de Tenerife, Spain
  • 2 Escuela de Doctorado y Estudios de Postgrado, Universidad de La Laguna, Tenerife, Spain
  • 3 Department of Molecular Genetics and Microbiology, Duke University Medical Center, Durham, NC 27710, USA
  • 4 Instituto de Tecnologías Biomédicas, Universidad de La Laguna, Tenerife, Spain
  • 5 Facultad de Ciencias de la Salud, Universidad Fernando Pessoa Canarias, Las Palmas de Gran Canaria, Spain
  • 6 Present address: BenchSci Analytics Inc., Toronto, Canada
  • 7 Present address: Genomic Integrity Unit, Danish Cancer Society Research Center, Copenhagen, Denmark

received: August 8, 2019 ; accepted: November 24, 2019 ; published: December 8, 2019 ;

https://doi.org/10.18632/aging.102573
How to Cite

Copyright © 2019 Ramos-Pérez et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Abstract

Topoisomerase II (Top2) removes topological linkages between replicated chromosomes. Top2 inhibition leads to mitotic catastrophe (MC) when cells unsuccessfully try to split their genetic material between the two daughter cells. Herein, we have characterized the fate of these daughter cells in the budding yeast. Clonogenic and microcolony experiments, in combination with vital and apoptotic stains, showed that 75% of daughter cells become senescent in the short term; they are unable to divide but remain alive. Decline in cell vitality then occurred, yet slowly, uncoordinatedly when comparing pairs of daughters, and independently of the cell death mediator Mca1/Yca1. Furthermore, we showed that senescence can be modulated by ploidy, suggesting that gross chromosome imbalances during segregation may account for this phenotype. Indeed, we found that diploid long-term survivors of the MC are prone to genomic imbalances such as trisomies, uniparental disomies and terminal loss of heterozygosity (LOH), the latter affecting the longest chromosome arms.

Abbreviations

MC: mitotic catastrophe; RCD: regulated cell death; ACD: accidental cell death; DSB: DNA double strand break; MB: methylene blue; PI: propidium iodide; ROS: reactive oxygen species; PS: phosphatidylserine; SNP: single nucleotide polymorphism; LOH: loss of heterozygosity; T-LOH: terminal LOH; I-LOH: interstitial LOH; UPD: uniparental disomy; RUPD: reciprocal UPD; rDNA: ribosomal RNA gene cluster.