Large Multimeric Assemblies of Nucleosome Assembly Protein and Histones Revealed by Small-angle X-ray Scattering and Electron Microscopy

Newman, Emily R.; Kneale, Geoff; Ravelli, Raimond B. G.; Krishnan, M.; Nejadasl, Fatemeh Karimi; Taylor, Ian A.; McGeehan, J.E.

doi:10.1074/jbc.m112.340422

Cited by 12 publications

(19 citation statements)

References 48 publications

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“…No such particles were observed in the absence of histones indicating that particle formation is dependent on H2A–H2B. Electron micrographs of yNap1c in complex with H2AΔ14–H2BΔ28 revealed monodisperse particles, about 15 nm in diameter, similar to those observed previously (Newman et al , 2012). Varying structural features indicated different orientations of the complex on the carbon support film (Fig 5A).…”

Section: Resultssupporting

confidence: 87%

“…Oligomerization of various Nap1 orthologs occurs in vivo and under physiological conditions in vitro (Ishimi et al , 1983, 1984; Fujii‐Nakata et al , 1992; Chang et al , 1997; Mosammaparast et al , 2002; McBryant & Peersen, 2004; Toth et al , 2005; Park et al , 2008; Noda et al , 2011; Newman et al , 2012). The biological role of Nap1 oligomerization has so far remained unclear.…”

Section: Resultsmentioning

confidence: 99%

“…In the presence of histones, yNap1 shows similar propensities to oligomerize (Toth et al , 2005; Noda et al , 2011). Xenopus laevis Nap1 in the presence of H2A–H2B or H3–H4 forms heterogeneous ring‐like particles in the 0.5–1.0 MDa size range as visualized by electron microscopy (Newman et al , 2012). The underlying compositional heterogeneity has made it difficult to ascertain the exact stoichiometry and to understand the structural basis and biological role of Nap1–histone oligomerization.…”

Section: Resultsmentioning

confidence: 99%

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Structural evidence for Nap1‐dependent H2A–H2B deposition and nucleosome assembly

et al. 2016

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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.

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Section: Resultssupporting

confidence: 87%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

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Structural evidence for Nap1‐dependent H2A–H2B deposition and nucleosome assembly

et al. 2016

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“…The reported cellular functions of Vps75 and Nap1 are with H3–H4 and H2A–H2B, respectively, although both chaperones are capable of binding both H2A–H2B and H3–H4 in vitro 54,55 . Vps75 and, to a lesser extent, Nap1 are capable of forming tetrameric ring-like assemblies 51,56 and they further oligomerize upon histone binding 51,52,57 . Tetramerization of Vps75 (FIG.…”

Section: Mechanistic Insights Into Chaperoning Histonesmentioning

confidence: 99%

Histone chaperone networks shaping chromatin function

Hammond

Strømme

Huang

et al. 2017

Nat Rev Mol Cell Biol

439

419

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The association of histones with specific chaperone complexes is important for their folding, oligomerization, post-translational modification, nuclear import, stability, assembly and genomic localization. In this way, the chaperoning of soluble histones is a key determinant of histone availability and fate, which affects all chromosomal processes, including gene expression, chromosome segregation and genome replication and repair. Here, we review the distinct structural and functional properties of the expanding network of histone chaperones. We emphasize how chaperones cooperate in the histone chaperone network and via co-chaperone complexes to match histone supply with demand, thereby promoting proper nucleosome assembly and maintaining epigenetic information by recycling modified histones evicted from chromatin.

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“…Xenopus laevis NAP1 (xNAP1) has also been found to form multimers with histones H2A‐H2B and H3‐H4 at a stoichiometry of one xNAP1 dimer to one histone fold dimer (Newman et al . ).…”

Section: Introductionmentioning

confidence: 97%

C‐terminal acidic domain of histone chaperone human NAP1 is an efficient binding assistant for histone H2A‐H2B, but not H3‐H4

et al. 2016

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Nucleosome assembly protein 1 (NAP1) binds both the (H3-H4) 2 tetramer and two H2A-H2B dimers, mediating their sequential deposition on DNA. NAP1 contains a C-terminal acidic domain (CTAD) and a core domain that promotes dimer formation. Here, we have investigated the roles of the core domain and CTAD of human NAP1 in binding to H2A-H2B and H3-H4 by isothermal calorimetry and native mass spectrometry and compared them with the roles of yeast NAP1. We show that the hNAP1 and yNAP1 dimers bind H2A-H2B by two different modes: a strong endothermic interaction and a weak exothermic interaction. A mutant hNAP1, but not yNAP1, dimer lacking CTAD loses the exothermic interaction and shows greatly reduced H2A-H2B binding activity. The isolated CTAD of hNAP1 binds H2A-H2B only exothermically with relatively stronger binding as compared with the exothermic interaction observed for the full-length hNAP1 dimer. Thus, the two CTADs in the hNAP1 dimer seem to provide binding assistance for the strong endothermic interaction of the core domain with H2A-H2B. By contrast, in the relatively weaker binding of hNAP1 to H3-H4 as compared with yNAP1, CTAD of hNAP1 has no significant role. To our knowledge, this is the first distinct role identified for the hNAP1 CTAD.

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Large Multimeric Assemblies of Nucleosome Assembly Protein and Histones Revealed by Small-angle X-ray Scattering and Electron Microscopy

Cited by 12 publications

References 48 publications

Structural evidence for Nap1‐dependent H2A–H2B deposition and nucleosome assembly

Structural evidence for Nap1‐dependent H2A–H2B deposition and nucleosome assembly

Histone chaperone networks shaping chromatin function

C‐terminal acidic domain of histone chaperone human NAP1 is an efficient binding assistant for histone H2A‐H2B, but not H3‐H4

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