An In Vitro Assay for Monitoring Topological DNA Entrapment by the Chromosomal Cohesin Complex

Methods Mol Biol. 2017:1515:23-35. doi: 10.1007/978-1-4939-6545-8_2.

Abstract

The cohesin complex is involved in a broad range of chromosomal biology, including DNA repair, gene transcription as well as sister chromatid cohesion. Cohesin is a large, ring-shaped protein complex and is thought to entrap DNA molecules inside of its ring. The unique DNA association is central to cohesin function and requires its ATPase and another heterodimer complex called the cohesin loader. Here we describe the biochemical reconstitution of topological cohesin loading onto DNA using the purified fission yeast cohesin proteins.

Keywords: Biochemical reconstitution; Chromosome segregation; Cohesin; Sister chromatid cohesion; Topological DNA entrapment.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphatases / genetics
  • Cell Cycle Proteins / genetics
  • Cell Cycle Proteins / isolation & purification*
  • Chromatids / genetics
  • Chromosomal Proteins, Non-Histone / genetics
  • Chromosomal Proteins, Non-Histone / isolation & purification*
  • Chromosome Segregation / genetics
  • Cohesins
  • DNA / genetics*
  • DNA Repair / genetics
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / isolation & purification
  • Molecular Biology / methods*
  • Saccharomyces cerevisiae Proteins / genetics
  • Saccharomyces cerevisiae Proteins / isolation & purification*
  • Sister Chromatid Exchange / genetics

Substances

  • Cell Cycle Proteins
  • Chromosomal Proteins, Non-Histone
  • DNA-Binding Proteins
  • Saccharomyces cerevisiae Proteins
  • DNA
  • Adenosine Triphosphatases