| First Author | Hajarnis SS | Year | 2015 |
| Journal | J Biol Chem | Volume | 290 |
| Issue | 41 | Pages | 24793-805 |
| PubMed ID | 26292219 | Mgi Jnum | J:340233 |
| Mgi Id | MGI:6838747 | Doi | 10.1074/jbc.M115.670646 |
| Citation | Hajarnis SS, et al. (2015) Transcription Factor Hepatocyte Nuclear Factor-1beta (HNF-1beta) Regulates MicroRNA-200 Expression through a Long Noncoding RNA. J Biol Chem 290(41):24793-805 |
| abstractText | The transcription factor hepatocyte nuclear factor-1beta (HNF-1beta) regulates tissue-specific gene expression in the kidney and other epithelial organs. Mutations of HNF-1beta produce kidney cysts, and previous studies have shown that HNF-1beta 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-1beta 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-1beta. HNF-1beta binds to two evolutionarily conserved sites located 28 kb upstream to miR-200b. Luciferase reporter assays showed that the HNF-1beta binding sites were located within a promoter that was active in renal epithelial cells. Mutations of the HNF-1beta 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-1beta knock-out mice and renal epithelial cells expressing dominant-negative mutant HNF-1beta. The expression of miR-200 targets, Zeb2 and Pkd1, was increased in HNF-1beta knock-out kidneys and in cells expressing mutant HNF-1beta. Overexpression of miR-200 decreased the expression of Zeb2 and Pkd1 in HNF-1beta mutant cells. These studies reveal a novel pathway whereby HNF-1beta directly contributes to the control of miRNAs that are involved in epithelial-mesenchymal transition and cystic kidney disease. |
