The core histone N-terminal domains are required for multiple rounds of catalytic chromatin remodeling by the SWI/SNF and RSC complexes

Biochemistry. 1999 Feb 23;38(8):2514-22. doi: 10.1021/bi982109d.

Abstract

SWI/SNF and RSC are large, distinct multi-subunit complexes that use the energy of ATP hydrolysis to disrupt nucleosome structure, facilitating the binding of transcription factors or restriction enzymes to nucleosomes [Cote, J., Quinn, J., Workman, J. L., and Peterson, C. L. (1994) Science 265, 53-60 (1); Lorch, Y., Cairns, B. R., Zhang, M., and Kornberg, R. D. (1998) Cell 94, 29-34 (2)]. Here we have used a quantitative assay to measure the activities of these ATP-dependent chromatin remodeling complexes using nucleosomal arrays reconstituted with hypoacetylated, hyperacetylated, or partially trypsinized histones. This assay is based on measuring the kinetics of restriction enzyme digestion of a site located within the central nucleosome of a positioned 11-mer array [Logie, C., and Peterson, C. L. (1997) EMBO J. 16, 6772-6782 (3)]. We find that the DNA-stimulated ATPase activities of SWI/SNF and RSC are not altered by the absence of the histone N-termini. Furthermore, ATP-dependent nucleosome remodeling is also equivalent on all three substrate arrays under reaction conditions where the concentrations of nucleosomal array and either SWI/SNF or RSC are equivalent. However, SWI/SNF and RSC cannot catalytically remodel multiple nucleosomal arrays in the absence of the histone termini, and this catalytic activity of SWI/SNF is decreased by histone hyperacetylation. These results indicate that the histone termini are important for SWI/SNF and RSC function; and, furthermore, our data defines a step in the remodeling cycle where the core histone termini exert their influence. This step appears to be after remodeling, but prior to intermolecular transfer of the remodelers to new arrays.

Publication types

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

MeSH terms

  • Acetylation
  • Adenosine Triphosphatases / metabolism
  • Catalysis
  • Chromosomal Proteins, Non-Histone
  • DNA / physiology
  • DNA-Binding Proteins / chemistry
  • DNA-Binding Proteins / metabolism*
  • Deoxyribonucleases, Type II Site-Specific / metabolism
  • Enzyme Activation
  • Fungal Proteins / chemistry
  • Fungal Proteins / metabolism*
  • Histones / chemistry
  • Histones / metabolism
  • Histones / physiology*
  • Kinetics
  • Macromolecular Substances
  • Models, Biological
  • Nucleosomes / chemistry
  • Nucleosomes / enzymology
  • Nucleosomes / metabolism
  • Peptide Fragments / chemistry
  • Peptide Fragments / metabolism
  • Peptide Fragments / physiology*
  • Protein Binding
  • Protein Folding
  • Protein Structure, Tertiary
  • Saccharomyces cerevisiae
  • Saccharomyces cerevisiae Proteins*
  • Salts
  • Substrate Specificity
  • Transcription Factors / chemistry
  • Transcription Factors / metabolism*

Substances

  • Chromosomal Proteins, Non-Histone
  • DNA-Binding Proteins
  • Fungal Proteins
  • Histones
  • Macromolecular Substances
  • Nucleosomes
  • Peptide Fragments
  • RSC complex, S cerevisiae
  • SWI1 protein, S cerevisiae
  • Saccharomyces cerevisiae Proteins
  • Salts
  • Transcription Factors
  • DNA
  • Deoxyribonucleases, Type II Site-Specific
  • GTYRAC-specific type II deoxyribonucleases
  • Adenosine Triphosphatases