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Monitoring the DNA Topoisomerase II Checkpoint in Saccharomyces cerevisiae

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DNA Topoisomerases

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1703))

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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 which 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.

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Authors and Affiliations

  1. Department of Genetics, Cell Biology & Development, University of Minnesota, 420 Washington Ave SE, Minneapolis, MN, 55455, USA

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

  2. St. John’s University, 2945 Abbey Plaza, Collegeville, MN, 56321, USA

    Katherine Furniss

  3. Biotechnology R&D, Cargill, Inc., 14800 28th Ave N, Plymouth, MN, 55447, USA

    Amit C. J. Vas

Authors
  1. Katherine Furniss
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  2. Amit C. J. Vas
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  3. Andrew B. Lane
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  4. Duncan J. Clarke
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Corresponding author

Correspondence to Duncan J. Clarke .

Editor information

Editors and Affiliations

  1. Département de microbiologie, infectiologie et immunologie, Université de Montréal, Montréal, Québec, Canada

    Marc Drolet

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Furniss, K., Vas, A.C.J., Lane, A.B., Clarke, D.J. (2018). Monitoring the DNA Topoisomerase II Checkpoint in Saccharomyces cerevisiae . In: Drolet, M. (eds) DNA Topoisomerases. Methods in Molecular Biology, vol 1703. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7459-7_16

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  • DOI: https://doi.org/10.1007/978-1-4939-7459-7_16

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7458-0

  • Online ISBN: 978-1-4939-7459-7

  • eBook Packages: Springer Protocols

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