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Comparative study of DNase I sensitivity at the X-linked humanHPRT locus

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Somatic Cell and Molecular Genetics

Abstract

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 HPRTgene 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 HPRTgene 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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Authors and Affiliations

  1. Department of Biochemistry, Baylor College of Medicine, 77030, Houston, Texas

    Dong Lin & A. Craig Chinault

  2. Institute for Molecular Genetics, Baylor College of Medicine, 77030, Houston, Texas

    A. Craig Chinault

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  1. Dong Lin
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  2. A. Craig Chinault
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Lin, D., Craig Chinault, A. Comparative study of DNase I sensitivity at the X-linked humanHPRT locus. Somat Cell Mol Genet 14, 261–272 (1988). https://doi.org/10.1007/BF01534587

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  • DOI: https://doi.org/10.1007/BF01534587

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