title: Unique yeast histone sequences influence octamer and nucleosome stability.
abstract: Yeast nucleosomes are known to be intrinsically less stable than those from higher eukaryotes. This difference presents significant challenges for the production of yeast nucleosome core particles (NCPs) and chromatin for in vitro analyses. Using recombinant yeast, human, and chimeric histone proteins, we demonstrate that three divergent amino acids in histone H3 (Q120 K121 K125 ) are responsible for the poor reconstitution of yeast histones into octamers. This QKK motif is only found in Fungi, and is located at the nucleosome dyad axis. Yeast-to-human changes at these positions render yeast histones amenable to well-established octamer reconstitution and salt dialysis methods for generating nucleosomal and longer chromatin templates. By contrast, the most divergent yeast core histones, H2A and H2B, affect the biophysical properties of NCP but not their stability. An evolutionary analysis of H3 sequences shows that a gradual divergence in H3 sequences occurred in Fungi to yield QKK in budding yeast. This likely facilitates the highly euchromatic nature of yeast genomes. Our results provide an explanation for the long recognized difference in yeast nucleosome stability and they offer a simple method to generate yeast chromatin templates for in vitro studies.
authors.affiliation: Department of Biochemistry and Microbiology, University of Victoria, British Columbia, Canada.
authors.forename: Andrew
authors.initials: A
authors.lastname: Leung
authors.affiliation: Department of Biochemistry and Microbiology, University of Victoria, British Columbia, Canada.
authors.forename: Manjinder
authors.initials: M
authors.lastname: Cheema
authors.affiliation: Chromatin Structure and Evolution (Chromevol) Group, Department of Biological Sciences, Florida International University, North Miami, FL, USA.
authors.forename: Rodrigo
authors.initials: R
authors.lastname: González-Romero
authors.affiliation: Chromatin Structure and Evolution (Chromevol) Group, Department of Biological Sciences, Florida International University, North Miami, FL, USA.
authors.forename: Jose M
authors.initials: JM
authors.lastname: Eirin-Lopez
authors.affiliation: Department of Biochemistry and Microbiology, University of Victoria, British Columbia, Canada.
authors.forename: Juan
authors.initials: J
authors.lastname: Ausió
authors.affiliation: Department of Biochemistry and Microbiology, University of Victoria, British Columbia, Canada.
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Nelson
mesh_terms: D000596:Amino Acids / Q000737:chemistry / Q000235:genetics; D002843:Chromatin / Q000737:chemistry / Q000235:genetics; D019143:Evolution, Molecular; D016681:Genome, Fungal; D006657:Histones / Q000737:chemistry / Q000235:genetics; D006801:Humans; D009707:Nucleosomes / Q000737:chemistry / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000235:genetics
chemical_list: D000596:Amino Acids; D002843:Chromatin; D006657:Histones; D009707:Nucleosomes; D011993:Recombinant Fusion Proteins
keywords: chromatin; nucleosome; yeast