title: Mineralocorticoid receptor antagonizes Dot1a-Af9 complex to increase αENaC transcription.
abstract: Aldosterone is a major regulator of Na(+) absorption and acts by activating the mineralocorticoid receptor (MR) to stimulate the epithelial Na(+) channel (ENaC). MR(-/-) mice exhibited pseudohypoaldosteronism type 1 (hyponatremia, hyperkalemia, salt wasting, and high levels of aldosterone) and died around postnatal day 10. However, if and how MR regulates ENaC transcription remain incompletely understood. Our earlier work demonstrated that aldosterone activates αENaC transcription by reducing expression of Dot1a and Af9 and by impairing Dot1a-Af9 interaction. Most recently, we reported identification of a major Af9 binding site in the αENaC promoter and upregulation of αENaC mRNA expression in mouse kidneys lacking Dot1a. Despite these findings, the putative antagonism between the MR/aldosterone and Dot1a-Af9 complexes has never been addressed. The molecular defects leading to PHA-1 in MR(-/-) mice remain elusive. Here, we report that MR competes with Dot1a to bind Af9. MR/aldosterone and Dot1a-Af9 complexes mutually counterbalance ENaC mRNA expression in inner medullary collecting duct 3 (IMCD3) cells. Real-time RT-quantitative PCR revealed that 5-day-old MR(-/-) vs. MR(+/+) mice had significantly lower αENaC mRNA levels. This change was associated with an increased Af9 binding and H3 K79 hypermethylation in the αENaC promoter. Therefore, this study identified MR as a novel binding partner and regulator of Af9 and a novel mechanism coupling MR-mediated activation with relief of Dot1a-Af9-mediated repression via MR-Af9 interaction. Impaired ENaC expression due to failure to inhibit Dot1a-Af9 may play an important role in the early stages of PHA-1 (before postnatal day 8) in MR(-/-) mice.
authors.affiliation: Dept. of Internal Medicine, Univ. of Texas Medical School at Houston, 6431 Fannin, MSB 5.135, Houston, TX 77030. wenzheng.zhang@uth.tmc.edu.
authors.forename: Xi
authors.initials: X
authors.lastname: Zhang
authors.forename: Qiaoling
authors.initials: Q
authors.lastname: Zhou
authors.forename: Lihe
authors.initials: L
authors.lastname: Chen
authors.forename: Stefan
authors.initials: S
authors.lastname: Berger
authors.forename: Hongyu
authors.initials: H
authors.lastname: Wu
authors.forename: Zhou
authors.initials: Z
authors.lastname: Xiao
authors.forename: David
authors.initials: D
authors.lastname: Pearce
authors.forename: Xiaodong
authors.initials: X
authors.lastname: Zhou
authors.forename: Wenzheng
authors.initials: W
authors.lastname: Zhang
mesh_terms: D000818:Animals; D001667:Binding, Competitive; D002460:Cell Line; D019175:DNA Methylation; D004195:Disease Models, Animal; D053503:Epithelial Sodium Channels / Q000235:genetics / Q000378:metabolism*; D020022:Genetic Predisposition to Disease; D011495:Histone-Lysine N-Methyltransferase; D006801:Humans; D007685:Kidney Tubules, Collecting / Q000201:enzymology*; D008780:Methyltransferases / Q000378:metabolism*; D051379:Mice; D018345:Mice, Knockout; D009687:Nuclear Proteins / Q000378:metabolism*; D010641:Phenotype; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D011546:Pseudohypoaldosteronism / Q000201:enzymology* / Q000235:genetics; D012333:RNA, Messenger / Q000378:metabolism; D018161:Receptors, Mineralocorticoid / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D014158:Transcription, Genetic; D014162:Transfection; D015854:Up-Regulation
chemical_list: D053503:Epithelial Sodium Channels; C489902:Mllt3 protein, mouse; D009687:Nuclear Proteins; D012333:RNA, Messenger; D018161:Receptors, Mineralocorticoid; C507047:Scnn1a protein, mouse; C481527:Dot1l protein, mouse; D008780:Methyltransferases; D011495:Histone-Lysine N-Methyltransferase
keywords: Af9; Dot1a; aldosterone; mineralocorticoid receptor; pseudohypoaldosteronism type 1
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