Aqp5 Is a New Transcriptional Target of Dot1a and a Regulator of Aqp2

Aqp5 Is a New Transcriptional Target of Dot1a and a Regulator of Aqp2

Dot1l encodes histone H3 K79 methyltransferase Dot1a. Mice with Dot1l deficiency in renal Aqp2-expressing cells (Dot1lAC) develop polyuria by unknown mechanisms. Here, we report that Aqp5 links Dot1l deletion to polyuria through Aqp2. cDNA array analysis revealed and real-time RT-qPCR validated Aqp...

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Main Authors: Wu, Hongyu, Chen, Lihe, Zhang, Xi, Zhou, Qiaoling, Li, Ju-Mei, Berger, Stefan, Borok, Zea, Zhou, Beiyun, Xiao, Zhou, Yin, Hongling, Liu, Mingyao, Wang, Ying, Jin, Jianping, Blackburn, Michael R., Xia, Yang, Zhang, Wenzheng
Format: Online
Language:English
Published: Public Library of Science 2013
Online Access:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3542343/
id pubmed-3542343
recordtype oai_dc
spelling pubmed-35423432013-01-16 Aqp5 Is a New Transcriptional Target of Dot1a and a Regulator of Aqp2 Wu, Hongyu Chen, Lihe Zhang, Xi Zhou, Qiaoling Li, Ju-Mei Berger, Stefan Borok, Zea Zhou, Beiyun Xiao, Zhou Yin, Hongling Liu, Mingyao Wang, Ying Jin, Jianping Blackburn, Michael R. Xia, Yang Zhang, Wenzheng Research Article Dot1l encodes histone H3 K79 methyltransferase Dot1a. Mice with Dot1l deficiency in renal Aqp2-expressing cells (Dot1lAC) develop polyuria by unknown mechanisms. Here, we report that Aqp5 links Dot1l deletion to polyuria through Aqp2. cDNA array analysis revealed and real-time RT-qPCR validated Aqp5 as the most upregulated gene in Dot1lAC vs. control mice. Aqp5 protein is barely detectable in controls, but robustly expressed in the Dot1lAC kidneys, where it colocalizes with Aqp2. The upregulation of Aqp5 is coupled with reduced association of Dot1a and H3 dimethyl K79 with specific subregions in Aqp5 5′ flanking region in Dot1lAC vs. control mice. In vitro studies in IMCD3, MLE-15 and 293Tcells using multiple approaches including real-time RT-qPCR, luciferase reporter assay, cell surface biotinylation assay, colocalization, and co-immunoprecipitation uncovered that Dot1a represses Aqp5. Human AQP5 interacts with AQP2 and impairs its cell surface localization. The AQP5/AQP2 complex partially resides in the ER/Golgi. Consistently, AQP5 is expressed in none of 15 normal controls, but in all of 17 kidney biopsies from patients with diabetic nephropathy. In the patients with diabetic nephropathy, AQP5 colocalizes with AQP2 in the perinuclear region and AQP5 expression is associated with impaired cellular H3 dimethyl K79. Taken together, these data for the first time identify Aqp5 as a Dot1a potential transcriptional target, and an Aqp2 binding partner and regulator, and suggest that the upregulated Aqp5 may contribute to polyuria, possibly by impairing Aqp2 membrane localization, in Dot1lAC mice and in patients with diabetic nephropathy. Public Library of Science 2013-01-10 /pmc/articles/PMC3542343/ /pubmed/23326416 http://dx.doi.org/10.1371/journal.pone.0053342 Text en © 2013 Wu et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
repository_type Open Access Journal
institution_category Foreign Institution
institution US National Center for Biotechnology Information
building NCBI PubMed
collection Online Access
language English
format Online
author Wu, Hongyu
Chen, Lihe
Zhang, Xi
Zhou, Qiaoling
Li, Ju-Mei
Berger, Stefan
Borok, Zea
Zhou, Beiyun
Xiao, Zhou
Yin, Hongling
Liu, Mingyao
Wang, Ying
Jin, Jianping
Blackburn, Michael R.
Xia, Yang
Zhang, Wenzheng
spellingShingle Wu, Hongyu
Chen, Lihe
Zhang, Xi
Zhou, Qiaoling
Li, Ju-Mei
Berger, Stefan
Borok, Zea
Zhou, Beiyun
Xiao, Zhou
Yin, Hongling
Liu, Mingyao
Wang, Ying
Jin, Jianping
Blackburn, Michael R.
Xia, Yang
Zhang, Wenzheng
Aqp5 Is a New Transcriptional Target of Dot1a and a Regulator of Aqp2
author_facet Wu, Hongyu
Chen, Lihe
Zhang, Xi
Zhou, Qiaoling
Li, Ju-Mei
Berger, Stefan
Borok, Zea
Zhou, Beiyun
Xiao, Zhou
Yin, Hongling
Liu, Mingyao
Wang, Ying
Jin, Jianping
Blackburn, Michael R.
Xia, Yang
Zhang, Wenzheng
author_sort Wu, Hongyu
title Aqp5 Is a New Transcriptional Target of Dot1a and a Regulator of Aqp2
title_short Aqp5 Is a New Transcriptional Target of Dot1a and a Regulator of Aqp2
title_full Aqp5 Is a New Transcriptional Target of Dot1a and a Regulator of Aqp2
title_fullStr Aqp5 Is a New Transcriptional Target of Dot1a and a Regulator of Aqp2
title_full_unstemmed Aqp5 Is a New Transcriptional Target of Dot1a and a Regulator of Aqp2
title_sort aqp5 is a new transcriptional target of dot1a and a regulator of aqp2
description Dot1l encodes histone H3 K79 methyltransferase Dot1a. Mice with Dot1l deficiency in renal Aqp2-expressing cells (Dot1lAC) develop polyuria by unknown mechanisms. Here, we report that Aqp5 links Dot1l deletion to polyuria through Aqp2. cDNA array analysis revealed and real-time RT-qPCR validated Aqp5 as the most upregulated gene in Dot1lAC vs. control mice. Aqp5 protein is barely detectable in controls, but robustly expressed in the Dot1lAC kidneys, where it colocalizes with Aqp2. The upregulation of Aqp5 is coupled with reduced association of Dot1a and H3 dimethyl K79 with specific subregions in Aqp5 5′ flanking region in Dot1lAC vs. control mice. In vitro studies in IMCD3, MLE-15 and 293Tcells using multiple approaches including real-time RT-qPCR, luciferase reporter assay, cell surface biotinylation assay, colocalization, and co-immunoprecipitation uncovered that Dot1a represses Aqp5. Human AQP5 interacts with AQP2 and impairs its cell surface localization. The AQP5/AQP2 complex partially resides in the ER/Golgi. Consistently, AQP5 is expressed in none of 15 normal controls, but in all of 17 kidney biopsies from patients with diabetic nephropathy. In the patients with diabetic nephropathy, AQP5 colocalizes with AQP2 in the perinuclear region and AQP5 expression is associated with impaired cellular H3 dimethyl K79. Taken together, these data for the first time identify Aqp5 as a Dot1a potential transcriptional target, and an Aqp2 binding partner and regulator, and suggest that the upregulated Aqp5 may contribute to polyuria, possibly by impairing Aqp2 membrane localization, in Dot1lAC mice and in patients with diabetic nephropathy.
publisher Public Library of Science
publishDate 2013
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3542343/
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