nuclear receptor shp activates mir-206 expression via a cascade dual inhibitory mechanism核受体轴马力通过级联激活mir - 206表达双重抑制机制.pdfVIP

Nuclear Receptor SHP Activates miR-206 Expression via a Cascade Dual Inhibitory Mechanism Guisheng Song, Li Wang* Departments of Medicine and Oncological Sciences, Huntsman Cancer Institute, University of Utah School of Medicine, Salt Lake City, Utah, United States of America Abstract MicroRNAs play a critical role in many essential cellular functions in the mammalian species. However, limited information is available regarding the regulation of miRNAs gene transcription. Microarray profiling and real-time PCR analysis revealed a marked down-regulation of miR-206 in nuclear receptor SHP2/ 2 mice. To understand the regulatory function of SHP with regard to miR-206 gene expression, we determined the putative transcriptional initiation site of miR-206 and also its full length primary transcript using a database mining approach and RACE. We identified the transcription factor AP1 binding sites on the miR-206 promoter and further showed that AP1 (c-Jun and c-Fos) induced miR-206 promoter transactivity and expression which was repressed by YY1. ChIP analysis confirmed the physical association of AP1 (c-Jun) and YY1 with the endogenous miR-206 promoter. In addition, we also identified nuclear receptor ERRc (NR3B3) binding site on the YY1 promoter and showed that YY1 promoter was transactivated by ERRc, which was inhibited by SHP (NROB2). ChIP analysis confirmed the ERRc binding to the YY1 promoter. Forced expression of SHP and AP1 induced miR-206 expression while overexpression of ERRc and YY1 reduced its expression. The effects of AP1, ERRc, and YY1 on miR-206 expression were reversed by siRNA knockdown of each gene, respectively. Thus, we propose a novel cascade ‘‘dual inhibitory’’ mechanism governing miR-206 gene transcription by SHP: SHP inhibition of ERRc led to decreased YY1 expression and the de- repression of YY1 o