All four core histone N-termini contain sequences required for the repression of basal transcription in yeast.
Nucleosomes prevent the recognition of TATA promoter elements by the basal transcriptional machinery in the absence of induction. However, while Saccharomyces cerevisiae histones H3 and H4 contain N-terminal regions involved in the activation and repression of GAL1 and in the expression of heterochromatin-like regions, the sequences involved in repressing basal transcription have not yet been identified. Here, we describe the mapping of new N-terminal domains, in all four core histones (H2A, H2B, H3 and H4), required for the repression of basal, uninduced transcription. Basal transcription was monitored by the use of a GAL1 promoter-URA3 reporter construct whose uninduced activity can be detected through cellular sensitivity to the drug, 5-fluoroorotic acid. We have found for each histone that the N-terminal sequences repressing basal activity are in a short region adjacent to the structured alpha-helical core. Analysis of minichromosome DNA topology demonstrates that the basal domains are required for the proper folding of DNA around the chromosomal particle. Deletion of the basal domain at each histone significantly decreases plasmid superhelical density, which probably reflects a release of DNA from the constraints of the nucleosome into the linker region. This provides a means by which basal factors may recognize otherwise repressed regulatory elements.
Lenfant
F
F
Department of Biological Chemistry, UCLA School of Medicine and Molecular Biology Institute, University of California, Los Angeles, CA 90095, USA.
Mann
R K
RK
Thomsen
B
B
Ling
X
X
Grunstein
M
M
D000107:Acetylation; D001483:Base Sequence; D002843:Chromatin / Q000648:ultrastructure; D015966:Gene Expression Regulation, Fungal; D017930:Genes, Reporter; D006657:Histones / Q000235:genetics*; D008969:Molecular Sequence Data; D009707:Nucleosomes / Q000378:metabolism; D010957:Plasmids / Q000378:metabolism; D011401:Promoter Regions, Genetic; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D017384:Sequence Deletion; D016385:TATA Box; D016615:Telomere / Q000235:genetics; D014158:Transcription, Genetic
D002843:Chromatin; D006657:Histones; D009707:Nucleosomes