title: Structural evidence for Nap1-dependent H2A-H2B deposition and nucleosome assembly.
abstract: Nap1 is a histone chaperone involved in the nuclear import of H2A-H2B and nucleosome assembly. Here, we report the crystal structure of Nap1 bound to H2A-H2B together with in vitro and in vivo functional studies that elucidate the principles underlying Nap1-mediated H2A-H2B chaperoning and nucleosome assembly. A Nap1 dimer provides an acidic binding surface and asymmetrically engages a single H2A-H2B heterodimer. Oligomerization of the Nap1-H2A-H2B complex results in burial of surfaces required for deposition of H2A-H2B into nucleosomes. Chromatin immunoprecipitation-exonuclease (ChIP-exo) analysis shows that Nap1 is required for H2A-H2B deposition across the genome. Mutants that interfere with Nap1 oligomerization exhibit severe nucleosome assembly defects showing that oligomerization is essential for the chaperone function. These findings establish the molecular basis for Nap1-mediated H2A-H2B deposition and nucleosome assembly.
authors.affiliation: European Molecular Biology Laboratory, Grenoble, France Unit for Virus Host-Cell Interactions, Univ. Grenoble Alpes-EMBL-CNRS, Grenoble, France.
authors.forename: Carmen
authors.initials: C
authors.lastname: Aguilar-Gurrieri
authors.affiliation: European Molecular Biology Laboratory, Grenoble, France Unit for Virus Host-Cell Interactions, Univ. Grenoble Alpes-EMBL-CNRS, Grenoble, France.
authors.forename: Amédé
authors.initials: A
authors.lastname: Larabi
authors.affiliation: Center for Eukaryotic Gene Regulation, Department of Biochemistry and Molecular Biology, The Pennsylvania State University, University Park, PA, USA.
authors.forename: Vinesh
authors.initials: V
authors.lastname: Vinayachandran
authors.affiliation: Department of Chemistry, University of Oxford, Oxford, UK.
authors.forename: Nisha A
authors.initials: NA
authors.lastname: Patel
authors.affiliation: Department of Cell Biology, Southern Medical University, Guangzhou, China.
authors.forename: Kuangyu
authors.initials: K
authors.lastname: Yen
authors.affiliation: Center for Eukaryotic Gene Regulation, Department of Biochemistry and Molecular Biology, The Pennsylvania State University, University Park, PA, USA.
authors.forename: Rohit
authors.initials: R
authors.lastname: Reja
authors.affiliation: Department of Chemistry, University of Oxford, Oxford, UK.
authors.forename: Ima-O
authors.initials: IO
authors.lastname: Ebong
authors.affiliation: Unit for Virus Host-Cell Interactions, Univ. Grenoble Alpes-EMBL-CNRS, Grenoble, France Université Grenoble-Alpes, Grenoble, France Centre National de la Recherche Scientifique (CNRS) IBS, Grenoble, France CEA, IBS, Grenoble, France.
authors.forename: Guy
authors.initials: G
authors.lastname: Schoehn
authors.affiliation: Department of Chemistry, University of Oxford, Oxford, UK.
authors.forename: Carol V
authors.initials: CV
authors.lastname: Robinson
authors.affiliation: Center for Eukaryotic Gene Regulation, Department of Biochemistry and Molecular Biology, The Pennsylvania State University, University Park, PA, USA.
authors.forename: B Franklin
authors.initials: BF
authors.lastname: Pugh
authors.affiliation: European Molecular Biology Laboratory, Grenoble, France Unit for Virus Host-Cell Interactions, Univ. Grenoble Alpes-EMBL-CNRS, Grenoble, France panne@embl.fr.
authors.forename: Daniel
authors.identifier: 0000-0001-9158-5507
authors.initials: D
authors.lastname: Panne
mesh_terms: D047369:Chromatin Immunoprecipitation; D018360:Crystallography, X-Ray; D004252:DNA Mutational Analysis; D006657:Histones / Q000737:chemistry* / Q000378:metabolism*; D008958:Models, Molecular; D056490:Nucleosome Assembly Protein 1 / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D009707:Nucleosomes / Q000378:metabolism*; D011485:Protein Binding; D011487:Protein Conformation; D055503:Protein Multimerization; D012441:Saccharomyces cerevisiae / Q000737:chemistry / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*
chemical_list: D006657:Histones; C533556:NAP1 protein, S cerevisiae; D056490:Nucleosome Assembly Protein 1; D009707:Nucleosomes; D029701:Saccharomyces cerevisiae Proteins
keywords: H2A–H2B; Nap1; chromatin; histone chaperone; nucleosome assembly