title: Biochemical and Functional Assays of Human Cohesin-Releasing Factor Wapl.
abstract: During the cell cycle, duplicated sister chromatids become physically connected during S phase through a process called sister-chromatid cohesion. Cohesion is terminated during the metaphase-to-anaphase transition to trigger sister-chromatid segregation. The establishment and dissolution of cohesion are highly regulated by the cohesin complex and its multitude of regulators. In particular, the cohesin regulator Wapl promotes the release of cohesin from chromosomes during both interphase and mitosis. Here, we describe in vitro protein binding assays between Wapl and a cohesin subcomplex, and cellular assays in human cells that probe the functions of Wapl in cohesin release.
authors: Howard Hughes Medical Institute, Department of Pharmacology, University of Texas Southwestern Medical Center, 6001 Forest Park Road, Dallas, TX, 75390, USA.
authors: Ge
authors: G
authors: Zheng
authors: Howard Hughes Medical Institute, Department of Pharmacology, University of Texas Southwestern Medical Center, 6001 Forest Park Road, Dallas, TX, 75390, USA.
authors: Zhuqing
authors: Z
authors: Ouyang
authors: Howard Hughes Medical Institute, Department of Pharmacology, University of Texas Southwestern Medical Center, 6001 Forest Park Road, Dallas, TX, 75390, USA. hongtao.yu@utsouthwestern.edu.
authors: Hongtao
authors: H
authors: Yu
mesh_terms: D002352:Carrier Proteins / Q000235:genetics / Q000302:isolation & purification*; D018797:Cell Cycle Proteins / Q000235:genetics / Q000302:isolation & purification*; D002842:Chromatids / Q000235:genetics; D002868:Chromosomal Proteins, Non-Histone / Q000235:genetics / Q000302:isolation & purification*; D020090:Chromosome Segregation / Q000235:genetics*; D006801:Humans; D008938:Mitosis / Q000235:genetics; D008967:Molecular Biology / Q000379:methods*; D009687:Nuclear Proteins / Q000235:genetics / Q000302:isolation & purification*; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000302:isolation & purification*; D012441:Saccharomyces cerevisiae / Q000235:genetics; D000097722:Cohesins
chemical_list: D002352:Carrier Proteins; D018797:Cell Cycle Proteins; D002868:Chromosomal Proteins, Non-Histone; D009687:Nuclear Proteins; D011518:Proto-Oncogene Proteins; D000097722:Cohesins; C488092:WAPL protein, human
keywords: Cohesin; Flow cytometry; In vitro binding assay; Metaphase spreads; Protein purification; Sister-chromatid cohesion; Wapl