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      Reactivation of an inactive human X chromosome: evidence for X inactivation by DNA methylation.

      Science (New York, N.Y.)
      Animals, Azacitidine, pharmacology, Base Sequence, Cell Differentiation, DNA, metabolism, Female, Gene Expression Regulation, drug effects, Glucosephosphate Dehydrogenase, genetics, Humans, Hybrid Cells, physiology, Hypoxanthine Phosphoribosyltransferase, Methylation, Mice, Sex Chromosomes, X Chromosome

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          Abstract

          A mouse-human somatic cell hybrid clone, deficient in hypoxanthine-guanine phosphoribosyltransferase (HPRT) and containing a structurally normal inactive human X chromosome, was isolated. The hybrid cells were treated with 5-azacytidine and tested for the reactivation and expression of human X-linked genes. The frequency of HPRT-positives clones after 5-azacytidine treatment was 1000-fold greater than that observed in untreated hybrid cells. Fourteen independent HPRT-positive clones were isolated and analyzed for the expression of human X markers. Isoelectric focusing showed that the HPRT expressed in these clones is human. One of the 14 clones expressed human glucose-6-phosphate dehydrogenase and another expressed human phosphoglycerate kinase. Since 5-azacytidine treatment results in hypomethylation of DNA, DNA methylation may be a mechanism of human X chromosome inactivation.

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