An In Vitro Assay for Cdc20-Dependent Mitotic Anaphase-Promoting Complex Activity from Budding Yeast [chapter]

Scott C. Schuyler, Andrew W. Murray
2009 Msphere  
Cell cycle transitions are controlled, in part, by ubiquitin-dependent proteolysis. In mitosis, the metaphase to anaphase transition is governed by an E3 ubiquitin ligase called the cyclosome or Anaphase-Promoting Complex (APC), and a WD40-repeat protein co-factor called Cdc20. In vitro Cdc20-dependent APC (APC Cdc20 ) assays have been useful in the identification and validation of target substrates, and in the study of APC enzymology and regulation. Many aspects of the regulation of cell cycle
more » ... progression have been discovered in the budding yeast Saccharomyces cerevisiae, and proteins purified from this model organism have been employed in a wide variety of in vitro assays. Here we outline a quantitative in vitro mitotic APC Cdc20 assay that makes use of a highly active form of the APC that is purified from budding yeast cells arrested in mitosis.
doi:10.1007/978-1-60327-993-2_17 pmid:19475395 fatcat:scdgbfuie5ftjnqs35epdsnhyu