PMID: 2480966Apr 1, 1989Paper

Activation of the hprt gene on the inactive X chromosome in transformed diploid female Chinese hamster cells.

Journal of Cell Science
S G Grant, R G Worton

Abstract

Treatment with 5-azacytidine, a potent inhibitor of DNA methylation, was used to induce activation of the selectable hprt gene on the inactive X chromosome in a diploid female Chinese hamster cell line. The transformed, stably diploid cell line F3B was selected in media containing the lethal purine analogue 6-thioguanine, to generate a phenotypically HPRT- mutant, F3BT1, of presumed genotype hprt-/hprt(+), where (+) represents the presumably wild-type allele on the inactive X chromosome. Treatment of F3BT1 with 5-azacytidine resulted in phenotypic reversion to HPRT+ at a frequency greater than 10(-3). Similar treatment of 6-thioguanine-resistant control lines derived from male cells, or from CHO (which has no inactive X chromosome), had no effect on the frequency of phenotypic reversion, indicating that activation of the hprt(+) allele, rather than reversion of the hprt- is responsible. This conclusion is substantiated by documentation of the low mutagenic capacity of 5-azacytidine in this system. Proof that the hprt(+) allele can be activated by 5-azacytidine treatment was obtained in somatic cell hybrids in which hprt gene products from the active and inactive X chromosomes could be distinguished by isoelectric focusing. Our ...Continue Reading

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