Regulation of MicroRNAs by Brahma-related Gene 1 (Brg1) in Smooth Muscle Cells

• Published in: The Journal of biological chemistry Vol. 288; no. 9; pp. 6397 - 6408
• Main Authors: Chen, Meng, Herring, B.Paul
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.03.2013; American Society for Biochemistry and Molecular Biology
• Subjects: Animals; Brg1; Chromatin Assembly and Disassembly - physiology; Chromatin Regulation; DNA Helicases - genetics; DNA Helicases - metabolism; Gene Regulation; Gene Transcription; Humans; Mice; Mice, Knockout; MicroRNA; MicroRNAs - biosynthesis; MicroRNAs - genetics; Multiprotein Complexes - genetics; Multiprotein Complexes - metabolism; Myocardin; Myocytes, Smooth Muscle - cytology; Myocytes, Smooth Muscle - metabolism; Myogenesis; Nuclear Proteins - biosynthesis; Nuclear Proteins - genetics; Nuclear Proteins - metabolism; Serum Response Factor; Serum Response Factor - genetics; Serum Response Factor - metabolism; Smooth Muscle; Trans-Activators - biosynthesis; Trans-Activators - genetics; Transcription Factors - genetics; Transcription Factors - metabolism; Gene Transcription; Serum Response Factor; MicroRNA; Brg1; Myocardin; Smooth Muscle; Myogenesis; Chromatin Regulation
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.M112.409474

MicroRNAs are involved in phenotypic switching of smooth muscle cells (SMCs). Brg1-containing SWI/SNF chromatin-remodeling complexes also play an important role in controlling the phenotype of SMCs. We thus determined whether Brg1 influences the transcription of microRNAs in SMCs. Microarray and quantitative RT-PCR analysis of smooth muscle from mice harboring smooth muscle-specific deletion of Brg1 revealed altered expression of several microRNAs, including miRs-143/145 and miR-133. Ablation of Brg1 in SMCs in vitro either by expression of dominant negative Brg1 or Brg1 knock-out attenuated miRs-143/145 expression. Knockdown of serum response factor (SRF) in SMCs significantly reduced the expression levels of miRs-143/145 and miR-133, whereas knockdown of myocardin only attenuated miRs-143/145 expression. Myocardin induced expression of miRs-143/145 and miR-133a and increased SRF binding to these genes in 10T1/2 cells. This myocardin-mediated induction was attenuated by dominant negative Brg1. In Brg1-null SW13 cells, miRs-143/145 were dramatically induced by myocardin only in the presence of Brg1, whereas miR-133 was not induced by myocardin in a Brg1-dependent manner. Chromatin immunoprecipitation assays demonstrated that in the presence of Brg1, myocardin increased SRF binding to both the miRs-143/145 and miR-133a loci. Together, these data suggest a mechanism in which Brg1-containing SWI/SNF complexes are required for myocardin to induce expression of miRs-143/145 in smooth muscle cells. In contrast, miR-133 expression appears to be regulated by Brg1-containing chromatin remodeling complexes in a partially SRF-dependent, although largely myocardin-independent manner. SWI/SNF-mediated chromatin remodeling thus regulates the phenotype of smooth muscle by affecting expression of protein-coding genes and microRNAs. Background: It is unknown whether Brg1 regulates microRNA expression during smooth muscle differentiation. Results: Brg1, SRF, and myocardin are required for transcription of miRs-143/145. Brg1 and SRF together with other factors regulate transcription of miR-133. Conclusion: Brg1 interacts with distinct factors to regulate expression of microRNAs. Significance: Brg1-containing chromatin-remodeling complexes regulate expression of both protein coding and noncoding genes to control smooth muscle differentiation.