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Methods designed for the identification and characterization ofin vitro andin vivo chromatin assembly mutants inSaccharomyces cerevisiae

Biological Procedures Online volume 5, pages 162–169 (2003)Cite this article

Abstract

Assembly of DNA into chromatin allows for the formation of a barrier that protects naked DNA from protein and chemical agents geared to degrade or metabolize DNA. Chromatin assembly occurs whenever a length of DNA becomes exposed to the cellular elements, whether during DNA synthesis or repair. This report describes tools to study chromatin assembly in the model systemSaccharomyces cerevisiae. Modifications to anin vitro chromatin assembly assay are described that allowed a brute force screen of temperature sensitive (ts) yeast strains in order to identify chromatin assembly defective extracts. This screen yielded mutations in genes encoding two ubiquitin protein ligases (E3s):RSP5, and a subunit of the Anaphase Promoting Complex (APC),APC5. Additional modifications are described that allow for a rapid analysis and anin vivo characterization of yeast chromatin assembly mutants, as well as any other mutant of interest. Our analysis suggests that thein vitro andin vivo chromatin assembly assays are responsive to different cellular signals, including cell cycle cues that involve different molecular networks.

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Authors and Affiliations

  1. Department of Anatomy and Cell Biology, University of Saskatchewan, S7N 5E5, Saskatoon, Saskatchewan

    Troy A. A. Harkness, Charmaine Legrand & Ashley Lone

  2. Department of Medicine, Royal University Hospital, University of Saskatchewan, S7N 0W8, Saskatoon, Saskatchewan

    Terra G. Arnason

Authors
  1. Troy A. A. Harkness
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  2. Terra G. Arnason
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  3. Charmaine Legrand
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  4. Ashley Lone
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Correspondence to Troy A. A. Harkness.

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Published: July 3, 2003

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Harkness, T.A.A., Arnason, T.G., Legrand, C. et al. Methods designed for the identification and characterization ofin vitro andin vivo chromatin assembly mutants inSaccharomyces cerevisiae . Biol. Proced. Online 5, 162–169 (2003). https://doi.org/10.1251/bpo58

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  • DOI: https://doi.org/10.1251/bpo58

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Biological Procedures Online

ISSN: 1480-9222

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