Abstract
The chromatin architecture of eukaryotic gene promoters is generally characterized by a nucleosome-free region (NFR) flanked by at least one H2A.Z variant nucleosome. Computational predictions of nucleosome positions based on thermodynamic properties of DNA-histone interactions have met with limited success. Here we show that the action of the essential RSC remodeling complex in S. cerevisiae helps explain the discrepancy between theory and experiment. In RSC-depleted cells, NFRs shrink such that the average positions of flanking nucleosomes move toward predicted sites. Nucleosome positioning at distinct subsets of promoters additionally requires the essential Myb family proteins Abf1 and Reb1, whose binding sites are enriched in NFRs. In contrast, H2A.Z deposition is dispensable for nucleosome positioning. By regulating H2A.Z deposition using a steroid-inducible protein splicing strategy, we show that NFR establishment is necessary for H2A.Z deposition. These studies suggest an ordered pathway for the assembly of promoter chromatin architecture.
Publication types
-
Research Support, N.I.H., Extramural
MeSH terms
-
Binding Sites
-
Cell Cycle Proteins / genetics
-
Cell Cycle Proteins / metabolism
-
Chromatin / genetics
-
Chromatin / metabolism*
-
Chromatin Assembly and Disassembly*
-
DNA-Binding Proteins / genetics
-
DNA-Binding Proteins / metabolism
-
Gene Expression Regulation, Fungal
-
Genome, Fungal
-
Histones / genetics
-
Histones / metabolism*
-
Nuclear Proteins / genetics
-
Nuclear Proteins / metabolism
-
Nucleosomes / genetics
-
Nucleosomes / metabolism*
-
Promoter Regions, Genetic
-
Protein Binding
-
Regulatory Sequences, Nucleic Acid / genetics
-
Saccharomyces cerevisiae / genetics
-
Saccharomyces cerevisiae / metabolism*
-
Saccharomyces cerevisiae Proteins / genetics
-
Saccharomyces cerevisiae Proteins / metabolism*
-
Transcription Factors / genetics
-
Transcription Factors / metabolism
Substances
-
ABF1 protein, S cerevisiae
-
Cell Cycle Proteins
-
Chromatin
-
DNA-Binding Proteins
-
Histones
-
Htz1 protein, S cerevisiae
-
Nuclear Proteins
-
Nucleosomes
-
REB1 protein, S cerevisiae
-
RSC complex, S cerevisiae
-
Saccharomyces cerevisiae Proteins
-
Transcription Factors
-
STH1 protein, S cerevisiae