Methods designed for the identification and characterization ofin vitro andin vivo chromatin assembly mutants inSaccharomyces cerevisiaeReport as inadecuate




Methods designed for the identification and characterization ofin vitro andin vivo chromatin assembly mutants inSaccharomyces cerevisiae - Download this document for free, or read online. Document in PDF available to download.

Biological Procedures Online

, Volume 5, Issue 1, pp 162–169

Received: 14 May 2003Revised: 17 June 2003Accepted: 23 June 2003DOI: 10.1251-bpo58

Cite this article as: Harkness, T.A.A., Arnason, T.G., Legrand, C. et al. Biol. Proced. Online 2003 5: 162. doi:10.1251-bpo58

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.

Indexing termsSaccharomyces cerevisiae Histones Ubiquitin Published: July 3, 2003

Download to read the full article text



Author: Troy A. A. Harkness - Terra G. Arnason - Charmaine Legrand - Ashley Lone

Source: https://link.springer.com/







Related documents