Fine mapping of the chromatin structure of a cell cycle-regulated human H4 histone gene.
We have observed changes in the chromatin structure of a human histone gene promoter that may be functionally related to variations in transcription during the cell cycle. A detailed analysis of the chromatin structure of a cell cycle-dependent human H4 histone gene and its flanking sequences was performed using DNase I, S1 nuclease, and restriction endonucleases. This gene was previously shown to have a DNase I- and S1-sensitive site for which the boundaries varied with the cell cycle, and we have now precisely mapped these modifications. During S phase, the entire coding region of this gene and the 5'-flanking region up to approximately -600 base pairs are sensitive to both DNase I and S1, while during mitosis/G1, accessibility to these enzymes is greatly decreased in regions from -250 to -600 base pairs and downstream of +100 base pairs. DNase I- and S1-hypersensitive sites in the proximal promoter region (which contains two sites of protein-DNA interaction as well as sequence elements necessary for the correct initiation of transcription) are present throughout the cell cycle, as is an additional site sensitive to both DNase I and S1, located at -700 to -800 base pairs. Restriction enzyme analysis confirmed the general openness of the promoter region and relative insensitivity of the 3'-flanking region, while salt wash experiments indicated several discrete sites in the promoter that are candidates for regulatory interactions. The chromatin structure of the proximal promoter region of this H4 gene is different during early S phase when it is maximally transcribed, as indicated by the ability of a high salt wash to render this region inaccessible to the restriction enzyme MspI only at this time of the cell cycle.
Chrysogelos
S
S
College of Medicine, University of Florida, Gainesville 32610.
Pauli
U
U
Stein
G
G
Stein
J
J
D002453:Cell Cycle; D002843:Chromatin / Q000648:ultrastructure*; D005786:Gene Expression Regulation; D005796:Genes; D006367:HeLa Cells / Q000166:cytology / Q000378:metabolism; D006657:Histones / Q000235:genetics*; D006801:Humans; D009693:Nucleic Acid Hybridization; D015183:Restriction Mapping
D002843:Chromatin; D006657:Histones