title: Purification of Yeast Native Reagents for the Analysis of Chromatin Function-II: Multiprotein Complexes and Biochemical Assays.
abstract: Post-translational modifications of histones play essential roles in regulating chromatin structure and function. These are tightly regulated in vivo and there is an intricate cross-talk between different marks as they are recognized by specific reader modules present in a large number of nuclear factors. In order to precisely dissect these processes in vitro native reagents like purified chromatin and histone modifying/remodeling enzymes are required to more accurately reproduce physiological conditions. The vast majority of these enzymes need to be part of stable multiprotein complexes with cofactors enabling them to act on chromatin substrates and/or read specific histone marks. In the accompanying chapter, we have described the protocol for purification of native chromatin from yeast cells (Chapter 3 ). Here, we present the methods to obtain highly purified native chromatin modifying complexes from Saccharomyces cerevisiae, based on Tandem Affinity Purification (TAP). We also present possible applications and useful functional assays that can be performed using these yeast native reagents.
authors.affiliation: St-Patrick Research Group in Basic Oncology, Laval University Cancer Research Center, CHU de Québec Research Center- Oncology axis, Hôtel-Dieu de Québec, 9 McMahon Street, Quebec City, QC, Canada, G1R 3S3.|Institut Curie/CNRS, UMR3664, Paris, France.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Lacoste
authors.affiliation: St-Patrick Research Group in Basic Oncology, Laval University Cancer Research Center, CHU de Québec Research Center- Oncology axis, Hôtel-Dieu de Québec, 9 McMahon Street, Quebec City, QC, Canada, G1R 3S3.
authors.forename: Wajid
authors.initials: W
authors.lastname: Bhat
authors.affiliation: St-Patrick Research Group in Basic Oncology, Laval University Cancer Research Center, CHU de Québec Research Center- Oncology axis, Hôtel-Dieu de Québec, 9 McMahon Street, Quebec City, QC, Canada, G1R 3S3. jacques.cote@crhdq.ulaval.ca.
authors.forename: Jacques
authors.initials: J
authors.lastname: Côté
mesh_terms: D001681:Biological Assay / Q000379:methods*; D002843:Chromatin / Q000378:metabolism*; D006657:Histones / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism
chemical_list: D002843:Chromatin; D006657:Histones; D029701:Saccharomyces cerevisiae Proteins
keywords: Chromatin; HAT; HMT; Histones; Purification; TAP; Yeast