Nuclease sensitivity and DNA methylation in estrogen regulation of Xenopus laevis vitellogenin gene expression

Abstract

Estrogen activates transcription of the vitellogenin genes in livers of male Xenopus laevis. We have examined the conformation of the vitellogenin genes in chromatin and the methylation state of one vitellogenin gene during the process of estrogen stimulation and withdrawal. Sensitivity of the vitellogenin genes to DNase I digestion parallels transcription. The vitellogenin genes are insensitive to DNase I digestion in unstimulated liver cells, become more sensitive to DNase I digestion following estrogen activation of vitellogenin gene transcription, and are insensitive to DNase I digestion in liver cells withdrawn from estrogen. In contrast, the methylation state of nine potential methylation sites within the vitellogenin A1 gene is identical in red blood cells, unstimulated and withdrawn liver, estrogen-stimulated liver, and hepatocytes purified from estrogen-stimulated liver. Rapid transcription of the vitellogenin genes in estrogen-stimulated liver cells occurs with six of the nine methylation sites examined fully methylated.