Butyrate induces selective transcriptional activation of a hypomethylated embryonic globin gene in adult erythroid cells.
An animal model of hemoglobin switching has been developed in which anemic adult chickens are treated with 5-azacytidine and sodium butyrate or alpha-aminobutyric acid, thereby resulting in activation of the embryonic rho-globin gene in adult erythroid cells. In vitro nuclear runoff transcription assays using erythroid nuclei from treated birds show that the mechanism of activation of the rho-globin gene is transcriptional whereas no transcriptional activation of the embryonic epsilon-globin gene occurs. The action of 5-azacytidine appears to be as an inhibitor of DNA methylation because other S-phase active cytotoxic drugs, when substituted for 5-azacytidine, do not cause demethylation of the embryonic globin genes, nor do they allow transcriptional activation to occur. Embryonic rho-globin gene activation in this model is not due to selection of primitive erythroid cells since a subpopulation of primitive erythroid cells is not evident either morphologically or when cells are probed for embryonic and adult globin RNA by in situ hybridization. These studies show that demethylation by 5-azacytidine is a prerequisite but not sufficient cis-regulatory event for a high level of transcriptional activation of the embryonic rho-globin gene in adult erythroid cells in vivo. The possible basis for the selective transcriptional activation by sodium butyrate in this system is discussed.
Burns
L J
LJ
Department of Internal Medicine, University of Iowa College of Medicine, Iowa City 52242.
Glauber
J G
JG
Ginder
G D
GD
D000107:Acetylation; D000818:Animals; D001374:Azacitidine / Q000494:pharmacology*; D015152:Blotting, Northern; D002087:Butyrates / Q000494:pharmacology*; D002645:Chickens; D005786:Gene Expression Regulation / Q000187:drug effects*; D005914:Globins / Q000235:genetics*; D006657:Histones / Q000502:physiology; D008745:Methylation; D010659:Phenylhydrazines / Q000494:pharmacology; D012156:Reticulocytes / Q000502:physiology; D014158:Transcription, Genetic / Q000187:drug effects*; D015533:Transcriptional Activation
D002087:Butyrates; D006657:Histones; D010659:Phenylhydrazines; C030299:phenylhydrazine; D005914:Globins; D001374:Azacitidine