Dynamic chromatin alterations triggered by natural and synthetic activation domains.
The activation domains (ADs) of transcription activators recruit a multiplicity of enzymatic activities to gene promoters. The mechanisms by which such recruitment takes place are not well understood. Using chromatin immunoprecipitation, we demonstrate dynamic alterations in the abundance of histones H2A, H3, and H4 at promoters of genes regulated by the HSF and Gal4 activators of Saccharomyces cerevisiae. Transcriptional activation of these genes, particularly those regulated by HSF, is accompanied by a significant reduction in both acetylated and unacetylated histones at promoters and may involve the transient displacement of histone octamers. To gain insight into the function of ADs, we conducted a genetic screen to identify polypeptides that could substitute for the 340-residue C-terminal activator of HSF and rescue the temperature sensitivity caused by its deletion. We found that the ts(-) phenotype of HSF(1-493) could be complemented by peptides as short as 11 amino acids. Such peptides are enriched in acidic and hydrophobic residues, and exhibit both trans-activating and chromatin-modifying activities when fused to the Gal4 DNA-binding domain. We also demonstrate that a previously identified 14-amino acid histone H3-binding module of human CTF1/NF1, which is similar to synthetic ADs, can substitute for the HSF C-terminal activator in conferring temperature resistance and can mediate the modification of promoter chromatin structure. Possible mechanisms of AD function, including one involving direct interactions with histones, are discussed.
Erkine
Alexander M
AM
Department of Biochemistry and Molecular Biology, Louisiana State University Health Sciences Center, Shreveport 71130, USA. aerkin@lsuhsc.edu
Gross
David S
DS
D000595:Amino Acid Sequence; D001483:Base Sequence; D015152:Blotting, Northern; D002843:Chromatin / Q000737:chemistry / Q000378:metabolism*; D017931:DNA Primers; D006360:Heat-Shock Proteins / Q000235:genetics; D006657:Histones / Q000378:metabolism; D010957:Plasmids; D011401:Promoter Regions, Genetic; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism
D002843:Chromatin; D017931:DNA Primers; D006360:Heat-Shock Proteins; D006657:Histones; D014157:Transcription Factors