Aldosterone increases renal tubular Na؉ absorption in large part by increasing transcription of the epithelial Na ؉ channel ␣-subunit (␣-ENaC) expressed in the apical membrane of collecting duct principal cells. We recently reported that a complex containing the histone H3K79 methyltransferase disruptor of telomeric silencing-1 (Dot1) associates with and represses the ␣-ENaC promoter in mouse inner medullary collecting duct mIMCD3 cells, and that aldosterone acts to disrupt this complex and its inhibitory effects (Zhang, W., Xia, X., Reisenauer, M. R., Rieg, T., Lang, F., Kuhl, D., Vallon, V., and Kone, B. C. Epithelial Na ϩ channel (ENaC) 2 is composed of three genetically distinct subunits (␣, , and ␥), and is expressed in the apical membrane of salt-absorbing epithelia of kidney, colon, and lung (1). ENaC expressed in the distal nephron constitutes the rate-limiting step for renal Na ϩ and fluid reabsorption and therefore is essential for physiological control of blood pressure and electrolyte composition. Genetic disorders of ENaC activity, such as pseudohypoaldosteronism type 1 and Liddle syndrome, are characterized by hypotension and hypertension, respectively (2). The number of channels present at the plasma membrane is highly regulated through channel biosynthesis and delivery to the plasma membrane and by channel retrieval from the plasma membrane (4 -6). Of the three ENaC subunits, it is synthesis of ␣-ENaC subunits that appears to be rate-limiting for the formation of new ENaC multimers. Thus mechanisms controlling ␣-ENaC gene transcription are central to the understanding of channel biosynthesis.Aldosterone, a key regulator of epithelial Na ϩ absorption, induces transepithelial Na ϩ transport in the collecting duct in large part through activation of ␣-ENaC gene transcription. Aldosterone administration or secondary hyperaldosteronism caused by salt restriction enhances ␣-ENaC gene transcription without increasing  or ␥ subunit expression or altering ␣-ENaC mRNA turnover (3). The biological actions of aldosterone on target gene transcription have generally been believed to be mediated principally or exclusively through the mineralocorticoid receptor (MR), a member of the nuclear hormone receptor family. Aldosterone has both immediate (Ͻ3 h) effects on ENaC, attributed to increased trafficking or activity of ENaC in the apical membrane, and delayed actions (Ͼ3 h) that involve the synthesis of new ENaC subunits (1,2,15,18,33). A highly conserved imperfect glucocorticoid-response element (GRE) has been identified in the 5Ј-flanking regions of the human, mouse, and rat ␣-ENaC genes and found to be necessary for trans-activation (12,17,35). Although heterologous expression of human ␣-ENaC in murine M1 collecting duct cells suggests that increased ␣-ENaC expression alone may be insufficient to increase ENaC activity under basal conditions, increased ␣-ENaC expression does appear to be necessary for full aldosterone induction of ENaC function (4).In prior work, we identified in the mouse inner medullary c...