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Reactivation ofhprt on the inactive X chromosome with DNA demethylating agents

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Somatic Cell and Molecular Genetics

Abstract

The C86 line of female embryonal carcinoma cells contains one active and one inactive X chromosome. Following methylnitrosourea mutagenesis, a clone called C86AGM2 was isolated that carries a mutated hprtgene on the active X chromosome. This hprtm allele encodes an HPRT enzyme that has less than 1% normal enzyme activity, is thermolabile, and has an altered isoelectric point. Following treatment with drugs that demethylate DNA, the hprt+ gene from the inactive X chromosome in C86AGM2 cells became active as determined by the appearance of HPRT activity with the thermodenaturation and electrofocusing characteristics of the normal enzyme. No expression of this hprt+ gene occurred if C86AGM2 cells were induced to differentiate prior to DNA demethylation. Stable lines of C86AGM2 cells expressing both the hprtm and hprt+ genes did not inactivate either gene following differentiation.

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Authors and Affiliations

  1. Departments of Medicine and Biology, University of Ottawa, 451 Smyth Road, K1H 8M5, Ottawa, Ontario, Canada

    Amanda J. Hockey, Chaker N. Adra & Michael W. McBurney

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  1. Amanda J. Hockey
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  2. Chaker N. Adra
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  3. Michael W. McBurney
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Hockey, A.J., Adra, C.N. & McBurney, M.W. Reactivation ofhprt on the inactive X chromosome with DNA demethylating agents. Somat Cell Mol Genet 15, 421–434 (1989). https://doi.org/10.1007/BF01534893

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  • DOI: https://doi.org/10.1007/BF01534893

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