@article{d02cc8c7384c429eafd662dfed3985d3,
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.",
keywords = "H2A–H2B, Nap1, chromatin, histone chaperone, nucleosome assembly",
author = "Carmen Aguilar-Gurrieri and Am{\'e}d{\'e} Larabi and Vinesh Vinayachandran and Patel, {Nisha A.} and Kuangyu Yen and Rohit Reja and Ebong, {Ima O.} and Guy Schoehn and Robinson, {Carol V.} and Pugh, {B. Franklin} and Daniel Panne",
note = "Funding Information: We thank the staff of ESRF and EMBL-Grenoble for assistance and support in using beamlines ID14–4, 23–1, and 23–2. We thank F. Mietton and J. Govin for in vivo analysis. We thank K. W. Muir for critical reading of the manuscript. We thank K. T. Simons and K. Luger for providing yNap1 and histone expression plasmids, respectively. Atomic coordinates and structure factors of the reported crystal structure have been deposited in the Protein Data Bank under accession code 5G2E. This work used the platforms of the Grenoble Instruct center (ISBG; UMS 3518 CNRS–CEA–UJF–EMBL) with support from FRISBI (ANR–10–INSB–05–02) and GRAL (ANR–10–LABX–49–01) within the Grenoble Partnership for Structural Biology (PSB). The EM platform is supported by the FRM, IBISA, the R{\'e}gion Rh{\^o}ne-Alpes and the Fond Feder. This work was supported by the ANR Blanc Grant “EPISTRUCT” ANR–GUI–AAP–04 to DP, and NIH grant HG004160 to BFP. Publisher Copyright: {\textcopyright} 2016 The Authors. Published under the terms of the CC BY NC ND 4.0 license",
year = "2016",
month = jul,
day = "1",
doi = "10.15252/embj.201694105",
language = "English (US)",
volume = "35",
pages = "1465--1482",
journal = "EMBO Journal",
issn = "0261-4189",
publisher = "Nature Publishing Group",
number = "13",
}