title: Cell-cycle-coupled structural oscillation of centromeric nucleosomes in yeast.
abstract: The centromere is a specialized chromosomal structure that regulates chromosome segregation. Centromeres are marked by a histone H3 variant. In budding yeast, the histone H3 variant Cse4 is present in a single centromeric nucleosome. Experimental evidence supports several different models for the structure of centromeric nucleosomes. To investigate Cse4 copy number in live yeast, we developed a method coupling fluorescence correlation spectroscopy and calibrated imaging. We find that centromeric nucleosomes have one copy of Cse4 during most of the cell cycle, whereas two copies are detected at anaphase. The proposal of an anaphase-coupled structural change is supported by Cse4-Cse4 interactions, incorporation of Cse4, and the absence of Scm3 in anaphase. Nucleosome reconstitution and ChIP suggests both Cse4 structures contain H2A/H2B. The increase in Cse4 intensity and deposition at anaphase are also observed in Candida albicans. Our experimental evidence supports a cell-cycle-coupled oscillation of centromeric nucleosome structure in yeast.
authors: Stowers Institute for Medical Research, Kansas City, MO 64110, USA.
authors: Manjunatha
authors: M
authors: Shivaraju
authors: Jay R
authors: JR
authors: Unruh
authors: Brian D
authors: BD
authors: Slaughter
authors: Mark
authors: M
authors: Mattingly
authors: Judith
authors: J
authors: Berman
authors: Jennifer L
authors: JL
authors: Gerton
mesh_terms: D000705:Anaphase; D002176:Candida albicans / Q000737:chemistry / Q000166:cytology* / Q000378:metabolism; D002453:Cell Cycle; D002503:Centromere / Q000378:metabolism*; D002868:Chromosomal Proteins, Non-Histone / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism; D005656:Fungal Proteins / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000032:analysis; D028861:Nuclear Pore Complex Proteins / Q000378:metabolism; D056490:Nucleosome Assembly Protein 1 / Q000378:metabolism; D009707:Nucleosomes / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000737:chemistry / Q000166:cytology* / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism
chemical_list: C092746:CSE4 protein, S cerevisiae; D002868:Chromosomal Proteins, Non-Histone; C467729:Cse4 protein, Candida albicans; D004268:DNA-Binding Proteins; D005656:Fungal Proteins; C533556:NAP1 protein, S cerevisiae; C077727:NUP49 protein, S cerevisiae; D028861:Nuclear Pore Complex Proteins; D056490:Nucleosome Assembly Protein 1; D009707:Nucleosomes; D029701:Saccharomyces cerevisiae Proteins; C521684:Scm3 protein, S cerevisiae; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins