Preferential binding of the histone (H3-H4)2 tetramer by NAP1 is mediated by the amino-terminal histone tails

J Biol Chem. 2003 Nov 7;278(45):44574-83. doi: 10.1074/jbc.M305636200. Epub 2003 Aug 19.

Abstract

The yeast nucleosome assembly protein 1 (yNAP1) participates in many diverse activities, such as the assembly of newly synthesized DNA into chromatin and the rearrangement of nucleosomes during transcriptional activation. yNAP1 does not require ATP hydrolysis to perform these functions and is a valuable tool for in vitro chromatin assembly. Using recombinant histone complexes, we show that yNAP1 has a preference for binding the (H3-H4)2 tetramer over the (H2A-H2B) dimer. We find that the loss of the histone tails abrogates this preference for H3 and H4, and we demonstrate a direct interaction between yNAP1 and the amino-terminal tails of H3 and H4. yNAP1 binds to one histone fold domain, thus specifying the stoichiometry of the complexes formed with the histone dimer and tetramer. Finally, we provide evidence that the acidic carboxyl-terminal region of yNAP1, although dispensable for nucleosome assembly in vitro, contributes to binding via structure-independent electrostatic interactions. Our results are consistent with recent mechanistic investigations of NAP1 and expand our understanding of the histone chaperone family of assembly factors.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Binding Sites
  • Cell Cycle Proteins
  • Chemical Phenomena
  • Chemistry, Physical
  • Chromatography, Gel
  • Dimerization
  • Drosophila / chemistry
  • Drosophila Proteins
  • Electrophoresis, Polyacrylamide Gel
  • Gene Deletion
  • Glutathione Transferase / genetics
  • Histones / chemistry*
  • Histones / metabolism*
  • Models, Molecular
  • Mutagenesis
  • Nuclear Proteins
  • Nucleosome Assembly Protein 1
  • Peptide Fragments / chemistry
  • Peptide Fragments / metabolism*
  • Polymerase Chain Reaction
  • Protein Folding
  • Proteins / chemistry
  • Proteins / genetics
  • Proteins / metabolism*
  • Recombinant Fusion Proteins
  • Saccharomyces cerevisiae / chemistry
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae Proteins
  • Static Electricity
  • Structure-Activity Relationship
  • Xenopus laevis

Substances

  • Cell Cycle Proteins
  • Drosophila Proteins
  • Histones
  • NAP1 protein, S cerevisiae
  • Nap1 protein, Drosophila
  • Nuclear Proteins
  • Nucleosome Assembly Protein 1
  • Peptide Fragments
  • Proteins
  • Recombinant Fusion Proteins
  • Saccharomyces cerevisiae Proteins
  • Glutathione Transferase