Kohei Ueda1, Katsunori Fujiki2, Katsuhiko Shirahige2, Celso E Gomez-Sanchez3, Toshiro Fujita4, Masaomi Nangaku1, Miki Nagase5. 1. Department of Nephrology and Endocrinology, The University of Tokyo, Tokyo, Japan. 2. Research Center for Epigenetic Disease, Institute of Molecular and Cellular Biosciences, The University of Tokyo, Tokyo, Japan. 3. Endocrine Section, G.V. (Sonny) Montgomery VA Medical Center, MS, USA; Endocrinology, University of Mississippi Medical Center, MS, USA. 4. Division of Clinical Epigenetics, Research Center for Advanced Science and Technology, The University of Tokyo, Tokyo, Japan. 5. Department of Nephrology and Endocrinology, The University of Tokyo, Tokyo, Japan; Department of Anatomy and Life Structure, School of Medicine Juntendo University, Tokyo, Japan. Electronic address: mnagase-tky@umin.ac.jp.
Abstract
BACKGROUND AND OBJECTIVE: Mineralocorticoid receptor (MR) is a member of nuclear receptor family proteins and contributes to fluid homeostasis in the kidney. Although aldosterone-MR pathway induces several gene expressions in the kidney, it is often unclear whether the gene expressions are accompanied by direct regulations of MR through its binding to the regulatory region of each gene. The purpose of this study is to identify the direct target genes of MR in a murine distal convoluted tubular epithelial cell-line (mDCT). METHODS: We analyzed the DNA samples of mDCT cells overexpressing 3xFLAG-hMR after treatment with 10(-7)M aldosterone for 1h by chromatin immunoprecipitation with deep-sequence (ChIP-seq) and mRNA of the cell-line with treatment of 10(-7)M aldosterone for 3h by microarray. RESULTS: 3xFLAG-hMR overexpressed in mDCT cells accumulated in the nucleus in response to 10(-9)M aldosterone. Twenty-five genes were indicated as the candidate target genes of MR by ChIP-seq and microarray analyses. Five genes, Sgk1, Fkbp5, Rasl12, Tns1 and Tsc22d3 (Gilz), were validated as the direct target genes of MR by quantitative RT-qPCR and ChIP-qPCR. MR binding regions adjacent to Ctgf and Serpine1 were also validated. CONCLUSIONS: We, for the first time, captured the genome-wide distribution of MR in mDCT cells and, furthermore, identified five MR target genes in the cell-line. These results will contribute to further studies on the mechanisms of kidney diseases.
BACKGROUND AND OBJECTIVE:Mineralocorticoid receptor (MR) is a member of nuclear receptor family proteins and contributes to fluid homeostasis in the kidney. Although aldosterone-MR pathway induces several gene expressions in the kidney, it is often unclear whether the gene expressions are accompanied by direct regulations of MR through its binding to the regulatory region of each gene. The purpose of this study is to identify the direct target genes of MR in a murine distal convoluted tubular epithelial cell-line (mDCT). METHODS: We analyzed the DNA samples of mDCT cells overexpressing 3xFLAG-hMR after treatment with 10(-7)M aldosterone for 1h by chromatin immunoprecipitation with deep-sequence (ChIP-seq) and mRNA of the cell-line with treatment of 10(-7)M aldosterone for 3h by microarray. RESULTS: 3xFLAG-hMR overexpressed in mDCT cells accumulated in the nucleus in response to 10(-9)M aldosterone. Twenty-five genes were indicated as the candidate target genes of MR by ChIP-seq and microarray analyses. Five genes, Sgk1, Fkbp5, Rasl12, Tns1 and Tsc22d3 (Gilz), were validated as the direct target genes of MR by quantitative RT-qPCR and ChIP-qPCR. MR binding regions adjacent to Ctgf and Serpine1 were also validated. CONCLUSIONS: We, for the first time, captured the genome-wide distribution of MR in mDCT cells and, furthermore, identified five MR target genes in the cell-line. These results will contribute to further studies on the mechanisms of kidney diseases.
Authors: P Mundel; J Reiser; A Zúñiga Mejía Borja; H Pavenstädt; G R Davidson; W Kriz; R Zeller Journal: Exp Cell Res Date: 1997-10-10 Impact factor: 3.905
Authors: Matthew A Deardorff; Masashige Bando; Ryuichiro Nakato; Erwan Watrin; Takehiko Itoh; Masashi Minamino; Katsuya Saitoh; Makiko Komata; Yuki Katou; Dinah Clark; Kathryn E Cole; Elfride De Baere; Christophe Decroos; Nataliya Di Donato; Sarah Ernst; Lauren J Francey; Yolanda Gyftodimou; Kyotaro Hirashima; Melanie Hullings; Yuuichi Ishikawa; Christian Jaulin; Maninder Kaur; Tohru Kiyono; Patrick M Lombardi; Laura Magnaghi-Jaulin; Geert R Mortier; Naohito Nozaki; Michael B Petersen; Hiroyuki Seimiya; Victoria M Siu; Yutaka Suzuki; Kentaro Takagaki; Jonathan J Wilde; Patrick J Willems; Claude Prigent; Gabriele Gillessen-Kaesbach; David W Christianson; Frank J Kaiser; Laird G Jackson; Toru Hirota; Ian D Krantz; Katsuhiko Shirahige Journal: Nature Date: 2012-09-13 Impact factor: 49.962
Authors: Katelee Barrett Mueller; Qing Lu; Najwa N Mohammad; Victor Luu; Amy McCurley; Gordon H Williams; Gail K Adler; Richard H Karas; Iris Z Jaffe Journal: Endocrinology Date: 2014-07-22 Impact factor: 4.736
Authors: Rachel V Richardson; Emma J Batchen; Adrian J W Thomson; Rowan Darroch; Xinlu Pan; Eva A Rog-Zielinska; Wiktoria Wyrzykowska; Kathleen Scullion; Emad A S Al-Dujaili; Mary E Diaz; Carmel M Moran; Christopher J Kenyon; Gillian A Gray; Karen E Chapman Journal: J Endocrinol Date: 2017-01-05 Impact factor: 4.286
Authors: Caroline A Rivers; Mark F Rogers; Felicity E Stubbs; Becky L Conway-Campbell; Stafford L Lightman; John R Pooley Journal: Endocrinology Date: 2019-05-01 Impact factor: 4.736
Authors: Jessica R Ivy; Wilna Oosthuyzen; Theresa S Peltz; Amelia R Howarth; Robert W Hunter; Neeraj Dhaun; Emad A S Al-Dujaili; David J Webb; James W Dear; Peter W Flatman; Matthew A Bailey Journal: Hypertension Date: 2016-03-07 Impact factor: 10.190