Comparative study of DNase I sensitivity at the X-linked human HPRT locus.
To examine the association between chromatin structure and gene expression at the human hypoxanthine phosphoribosyltransferase (HPRT) locus, DNase I sensitivity of active and inactive genes was analyzed. In a set of human-hamster hybrid lines containing either an active or an inactive human X chromosome, or a derivative of the latter in which the HPRT gene was reactivated by 5-azacytidine treatment, only the promoter region of the gene was found to contain a hypersensitive domain, and its presence was strictly correlated with gene activity. An S1 nuclease-sensitive site was mapped upstream from the DNase I hypersensitive domain using supercoiled plasmids. The overall level of DNase I sensitivity in the interior of the HPRT gene was also assessed by comparing the degradation of polymorphic restriction fragments on active and inactive alleles in both polyclonal and monoclonal lines of female human cells. In these internally controlled experiments, the active X chromosome was found to be approximately twofold more susceptible to DNase I digestion than the inactive X chromosome.
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Department of Biochemistry, Baylor College of Medicine, Houston, Texas 77030.
Chinault
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D000818:Animals; D002874:Chromosome Mapping; D006224:Cricetinae; D004262:DNA Restriction Enzymes; D003850:Deoxyribonuclease I / Q000235:genetics*; D004303:Dosage Compensation, Genetic; D008040:Genetic Linkage; D005819:Genetic Markers; D006801:Humans; D006822:Hybrid Cells; D007041:Hypoxanthine Phosphoribosyltransferase / Q000235:genetics*; D014960:X Chromosome
D005819:Genetic Markers; D007041:Hypoxanthine Phosphoribosyltransferase; D004262:DNA Restriction Enzymes; D003850:Deoxyribonuclease I