Methyl binding domain protein 2 mediates γ-globin gene silencing in adult human βYAC transgenic mice

Abstract

The genes of the vertebrate β-globin locus undergo a switch in expression during erythroid development whereby embryonic/fetal genes of the cluster are sequentially silenced and adult genes are activated. We describe here a role for DNA methylation and MBD2 in the silencing of the human fetal γ-globin gene. The γ-globin gene is reactivated upon treatment with the DNA methyltransferase inhibitor 5-azacytidine in the context of a mouse containing the entire human β-globin locus as a yeast artificial chromosome (βYAC) transgene. To elucidate the mechanism through which DNA methylation represses the γ-globin gene in adult erythroid cells, βYAC/MBD2−/− mice were generated by breeding βYAC mice with MBD2−/− mice. Adult βYAC/MBD2−/− mice continue to express the γ-globin gene at a level commensurate with 5-azacytidine treatment, 10- to 20-fold over that observed with 1-acetyl-2-phenylhydrazine treatment alone. In addition, the level of γ-globin expression is consistently higher in MBD2−/− mice in 14.5- and 16.5-days postcoitus fetal liver erythroblasts suggesting a role for MBD2 in embryonic/fetal erythroid development. DNA methylation levels are modestly decreased in MBD2−/− mice. MBD2 does not bind to the γ-globin promoter region to maintain γ-globin silencing. Finally, treatment of MBD2-null mice with 5-azacytidine induces only a small, nonadditive induction of γ-globin mRNA, signifying that DNA methylation acts primarily through MBD2 to maintain γ-globin suppression in adult erythroid cells.