AF17 Competes With AF9 for Binding to DOT1A to up-Regulate Transcription of Epithelial NA+ Channel α
Document Type
Article
Publication Date
12-18-2009
Description
We previously reported that Dot1a*AF9 complex represses transcription of the epithelial Na+ channel subunit α (α-ENaC) gene in mouse inner medullary collecting duct mIMCD3 cells and mouse kidney. Aldosterone relieves this repression by down-regulating the complex through various mechanisms. Whether these mechanisms are sufficient and conserved in human cells or can be applied to other aldosterone-regulated genes remains largely unknown. Here we demonstrate that human embryonic kidney 293T cells express the three ENaC subunits and all of the ENaC transcriptional regulators examined. These cells respond to aldosterone and display benzamil-sensitive Na+ currents, as measured by whole-cell patch clamping. We also show that AF17 and AF9 competitively bind to the same domain of Dot1a in multiple assays and have antagonistic effects on expression of an α-ENaC promoter-luciferase construct. Overexpression of Dot1a or AF9 decreased mRNA expression of the ENaC subunits and their transcriptional regulators and reduced benzamil-sensitive Na+ currents. AF17 over-expression caused the opposite effects, accompanied by redirection of Dot1a from the nucleus to the cytoplasm and reduction in histone H3 K79 methylation. The nuclear export inhibitor leptomycin B blocked the effect of AF17 overexpression on H3 K79 hypomethylation. RNAi-mediated knockdown of AF17 yielded nuclear enrichment of Dot1a and histone H3 K79 hypermethylation. As with AF9, AF17 displays nuclear and cytoplasmic co-localization with Sgk1. Therefore, AF17 competes with AF9 to bind Dot1a, decreases Dot1a nuclear expression by possibly facilitating its nuclear export, and relieves Dot1a*AF9-mediated repression of α-ENaC and other target genes.
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This work is licensed under a Creative Commons Attribution 4.0 International License.
Citation Information
Reisenauer, Mary R.; Anderson, Marc; Huang, Le; Zhang, Zhijing; Zhou, Qiaoling; Kone, Bruce C.; Morris, Andrew P.; LeSage, Gene D.; Dryer, Stuart E.; and Zhang, Wenzheng. 2009. AF17 Competes With AF9 for Binding to DOT1A to up-Regulate Transcription of Epithelial NA+ Channel α. Journal of Biological Chemistry. Vol.284(51). 35659-35669. https://doi.org/10.1074/jbc.M109.038448 PMID: 19864429 ISSN: 0021-9258
Copyright Statement
© 2009 ASBMB. Currently published by Elsevier Inc; originally published by American Society for Biochemistry and Molecular Biology.
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