In vitro assembly of physiological cohesin/DNA complexes

Itay Onn, Douglas Koshland

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22 Scopus citations


Cohesin is a member of the Smc family of protein complexes that mediates higher-order chromosome structure by tethering different regions of chromatin. We present a new in vitro system that assembles cohesin-DNA complexes with in vivo properties. The assembly of these physiological salt-resistant complexes requires the cohesin holo-complex, its ability to bind ATP, the cohesin loader Scc2p and a closed DNA topology. Both the number of cohesin molecules bound to the DNA substrate and their distribution on the DNA substrate are limited. Cohesin and Scc2p bind preferentially to cohesin associated regions (CARs), DNA sequences with enriched cohesin binding in vivo. A subsequence of CARC1 promotes cohesin binding to neighboring sequences within CARC1. The enhancer-like function of this sequence is validated by in vivo deletion analysis. By demonstrating the physiological relevance of these in vitro assembled cohesin-DNA complexes, we establish our in vitro system as a powerful tool to elucidate the mechanism of cohesin and other Smc complexes.

Original languageEnglish
Pages (from-to)12198-12205
Number of pages8
JournalProceedings of the National Academy of Sciences of the United States of America
Issue number30
StatePublished - 26 Jul 2011
Externally publishedYes


  • Cohesion
  • DNA binding
  • Structural maintenance of chromosomes
  • Yeast


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