Abstract |
The transcription factor hepatocyte nuclear factor-1β (HNF-1β) regulates tissue-specific gene expression in the kidney and other epithelial organs. Mutations of HNF-1β produce kidney cysts, and previous studies have shown that HNF-1β regulates the transcription of cystic disease genes, including Pkd2 and Pkhd1. Here, we combined chromatin immunoprecipitation and next-generation sequencing (ChIP-Seq) with microarray analysis to identify microRNAs ( miRNAs) that are directly regulated by HNF-1β in renal epithelial cells. These studies identified members of the epithelial-specific miR-200 family (miR-200b/200a/429) as novel transcriptional targets of HNF-1β. HNF-1β binds to two evolutionarily conserved sites located 28 kb upstream to miR-200b. Luciferase reporter assays showed that the HNF-1β binding sites were located within a promoter that was active in renal epithelial cells. Mutations of the HNF-1β binding sites abolished promoter activity. RT-PCR analysis revealed that a long noncoding RNA ( lncRNA) is transcribed from the promoter and encodes the miR-200 cluster. Inhibition of the lncRNA with siRNAs decreased the levels of miR-200 but did not affect expression of the Ttll10 host gene. The expression of the lncRNA and miR-200 was decreased in kidneys from HNF-1β knock-out mice and renal epithelial cells expressing dominant-negative mutant HNF-1β. The expression of miR-200 targets, Zeb2 and Pkd1, was increased in HNF-1β knock-out kidneys and in cells expressing mutant HNF-1β. Overexpression of miR-200 decreased the expression of Zeb2 and Pkd1 in HNF-1β mutant cells. These studies reveal a novel pathway whereby HNF-1β directly contributes to the control of miRNAs that are involved in epithelial-mesenchymal transition and cystic kidney disease.
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Authors | Sachin S Hajarnis, Vishal Patel, Karam Aboudehen, Massimo Attanasio, Patricia Cobo-Stark, Marco Pontoglio, Peter Igarashi |
Journal | The Journal of biological chemistry
(J Biol Chem)
Vol. 290
Issue 41
Pg. 24793-805
(Oct 09 2015)
ISSN: 1083-351X [Electronic] United States |
PMID | 26292219
(Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)
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Copyright | © 2015 by The American Society for Biochemistry and Molecular Biology, Inc. |
Chemical References |
- Hnf1b protein, mouse
- Homeodomain Proteins
- MicroRNAs
- Mirn200 microRNA, mouse
- RNA, Long Noncoding
- Repressor Proteins
- TRPP Cation Channels
- ZEB2 protein, human
- Zinc Finger E-box Binding Homeobox 2
- polycystic kidney disease 1 protein
- Hepatocyte Nuclear Factor 1-beta
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Topics |
- Animals
- Base Sequence
- Epithelial Cells
(metabolism)
- Gene Expression Regulation
- Gene Knockout Techniques
- Genomics
- HeLa Cells
- Hepatocyte Nuclear Factor 1-beta
(deficiency, genetics, metabolism)
- Homeodomain Proteins
(genetics)
- Humans
- Kidney
(cytology)
- Mice
- MicroRNAs
(genetics)
- Mutation
- RNA, Long Noncoding
(genetics)
- Repressor Proteins
(genetics)
- TRPP Cation Channels
(genetics)
- Zinc Finger E-box Binding Homeobox 2
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