Genomic Nucleosome Organization Reconstituted with Pure Proteins

Nils Krietenstein, Megha Wal, Shinya Watanabe, Bongsoo Park, Craig L. Peterson, B. Franklin Pugh, Philipp Korber

Research output: Contribution to journalArticlepeer-review

Abstract

Chromatin remodelers regulate genes by organizing nucleosomes around promoters, but their individual contributions are obfuscated by the complex in vivo milieu of factor redundancy and indirect effects. Genome-wide reconstitution of promoter nucleosome organization with purified proteins resolves this problem and is therefore a critical goal. Here, we reconstitute four stages of nucleosome architecture using purified components: yeast genomic DNA, histones, sequence-specific Abf1/Reb1, and remodelers RSC, ISW2, INO80, and ISW1a. We identify direct, specific, and sufficient contributions that in vivo observations validate. First, RSC clears promoters by translating poly(dA:dT) into directional nucleosome removal. Second, partial redundancy is recapitulated where INO80 alone, or ISW2 at Abf1/Reb1sites, positions +1 nucleosomes. Third, INO80 and ISW2 each align downstream nucleosomal arrays. Fourth, ISW1a tightens the spacing to canonical repeat lengths. Such a minimal set of rules and proteins establishes core mechanisms by which promoter chromatin architecture arises through a blend of redundancy and specialization.

Original languageEnglish (US)
Pages (from-to)709-721.e12
JournalCell
Volume167
Issue number3
DOIs
StatePublished - Oct 20 2016

Keywords

  • Abf1
  • INO80
  • Isw
  • RSC
  • Reb1
  • Saccharomyces cerevisiae
  • chromatin
  • general regulatory factors (GRFs)
  • in vitro reconstitution
  • nucleosome positioning and remodeling

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Fingerprint

Dive into the research topics of 'Genomic Nucleosome Organization Reconstituted with Pure Proteins'. Together they form a unique fingerprint.

Cite this