Proximal and long-range alterations in chromatin structure surrounding the Chinese hamster dihydrofolate reductase promoter.
The chromatin structure of the dihydrofolate reductase (DHFR) gene was examined by DNA/protein cross-linking, chemical DNA methylation, and micrococcal nuclease digestion. The 5' promoter region of the gene displays two nucleosome-free zones (-550 to -300 and -150 to +100 bp relative to the ATG codon), each of which contains a number of micrococcal nuclease-hypersensitive sites. Regions upstream from the distal hypersensitive zone (-900 to -550 bp), downstream from the proximal hypersensitive zone (+100 to +400 bp), and between these two zones (-300 to -150 bp) appear either to be more than 80% histone-free or to contain histones whose globular domains have lost most of their contacts with DNA. Overall, a broad zone extending from -4300 to +4700 bp is altered relative to bulk chromatin, and within this region there are positioned nucleosomes and/or nucleosome-free zones in which the DNA appears to interact with a number of different non-histone proteins. By comparison, the chromatin in the 3' end of the gene (including the right end of the 5th intron and the 6th exon) contains randomly positioned nucleosomes, and its structure is intermediate between that of the 5' end of the gene and a downstream matrix attachment region that contains regularly organized chromatin. A 2.3 kb zone in the central part of the 5th intron reveals some features similar to the 5' end of the gene, suggesting a hitherto unrecognized functional role.
Engelhardt Institute of Molecular Biology, Moscow, Russia.
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Pemov
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Bavykin
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JL
Hamlin
D000818:Animals; D002460:Cell Line; D002843:Chromatin / Q000648:ultrastructure*; D006224:Cricetinae; D003412:Cricetulus; D003432:Cross-Linking Reagents; D004247:DNA / Q000737:chemistry / Q000648:ultrastructure; D015971:Gene Expression Regulation, Enzymologic; D006657:Histones / Q000737:chemistry; D066298:In Vitro Techniques; D008745:Methylation; D008836:Micrococcal Nuclease / Q000494:pharmacology; D009707:Nucleosomes / Q000648:ultrastructure; D011401:Promoter Regions, Genetic; D013463:Sulfuric Acid Esters / Q000737:chemistry; D013762:Tetrahydrofolate Dehydrogenase / Q000235:genetics*
D002843:Chromatin; D003432:Cross-Linking Reagents; D006657:Histones; D009707:Nucleosomes; D013463:Sulfuric Acid Esters; D004247:DNA; D013762:Tetrahydrofolate Dehydrogenase; D008836:Micrococcal Nuclease; C007482:dimethyl sulfate