Assaying topoisomerase II checkpoints in yeast.

Katherine Furniss, Amit C.J. Vas, Andrew Lane, Duncan J. Clarke

Research output: Contribution to journalArticle

2 Scopus citations

Abstract

Topoisomerase II activity is crucial to maintain genome stability through the removal of catenanes in the DNA formed during DNA replication and scaffolding the mitotic chromosome. Perturbed Topo II activity causes defects in chromosome segregation due to persistent catenations and aberrant DNA condensation during mitosis. Recently, novel top2 alleles in the yeast Saccharomyces cerevisiae revealed a checkpoint control that responds to perturbed Topo II activity. Described in this chapter are protocols for assaying the phenotypes seen in top2 mutants on a cell biological basis in live cells: activation of the Topo II checkpoint using spindle morphology, chromosome condensation using fluorescently labeled chromosomal loci, and cell cycle progression by flow cytometry. Further characterization of this novel checkpoint is warranted so that we can further our understanding of the cell cycle, genomic stability, and the possibility of identifying novel drug targets.

Original languageEnglish (US)
Pages (from-to)167-187
Number of pages21
JournalMethods in molecular biology (Clifton, N.J.)
Volume582
StatePublished - 2009

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