Analyzing mitotic chromosome structural defects after topoisomerase II inhibition or mutation

Juan F. Giménez-Abián, Andrew B. Lane, Duncan J. Clarke

Research output: Chapter in Book/Report/Conference proceedingChapter

1 Scopus citations

Abstract

For analyzing chromosome structural defects that result from topoisomerase II (topo II) dysfunction we have adapted classical cell cycle experiments, classical cytological techniques and the use of a potent topo II inhibitor (ICRF-193). In this chapter, we describe in detail the protocols used and we discuss the rational for our choice and for the adaptations applied. We clarify in which cell cycle stages each of the different chromosomal aberrations induced by inhibiting topo II takes place: lack of chromosome segregation, undercondensation, lack of sister chromatid resolution, and lack of chromosome individualization. We also put these observations into the context of the two topo II-dependent cell cycle checkpoints. In addition, we have devised a system to analyze phenotypes that result when topo II is mutated in human cells. This serves as an alternative strategy to the use of topo II inhibitors to perturb topo II function.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages191-215
Number of pages25
DOIs
StatePublished - 2018

Publication series

NameMethods in Molecular Biology
Volume1703
ISSN (Print)1064-3745

Keywords

  • Chromatid resolution
  • Chromosome individualization
  • Condensation
  • ICRF-193
  • Topo II checkpoint
  • Topoisomerase II

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