AML1 IS EXPRESSED IN SKELETAL-MUSCLE AND IS REGULATED BY INNERVATION

Although most skeletal muscle genes are expressed at similar levels in electrically active, innervated muscle and in electrically inactive, denervated muscle, a small number of genes, including those encoding t he acetylcholine receptor, N-CAM, and myogenin, are expressed at signi ficantly higher levels in denervated than in innervated muscle. The me chanisms that mediate electrical activity-dependent gene regulation ar e not understood, but these mechanisms are likely to be responsible, a t least in part, for the changes in muscle structure and function that accompany a decrease in myofiber electrical activity. To understand b ow muscle activity regulates muscle structure and function, we used a subtractive-hybridization and cloning strategy to identify and isolate genes that are expressed preferentially in innervated or denervated m uscle. One of the genes which we found to be regulated by electrical a ctivity is the recently discovered acute myeloid leukemia 1 (AML1) gen e. Disruption and translocation of the human AML1 gene are responsible for a form of acute myeloid leukemia. AML1 is a DNA-binding protein, but its normal function is not known and its expression and regulation in skeletal muscle were not previously appreciated. Because of its po tential role as a transcriptional mediator of electrical activity, we characterized expression of the AML1 gene in innervated, denervated, a nd developing skeletal muscle. We show that AML1 is expressed at low l evels in innervated skeletal muscle and at 50- to 100-fold-higher leve ls in denervated muscle. Four AML1 transcripts are expressed in denerv ated muscle, and the abundance of each transcript increases after dene rvation. We transfected C2 muscle cells with an expression vector enco ding AML1, tagged with an epitope from hemagglutinin, and we show that AML1 is a nuclear protein in muscle. AML1 dimerizes with core-binding factor beta (CBF beta), and we show that CBF beta is expressed at hig h levels in both innervated and denervated skeletal muscle. PEBP2 alph a, which is structurally related to AML1 and which also dimerizes with CBF beta, is expressed at low levels in skeletal muscle and is up-reg ulated only weakly by denervation. These results are consistent with t he idea that AML1 may have a role in regulating gene expression in ske letal muscle.