Analyzing mRNA expression identifies Smad3 as a microRNA-140 target regulated only at protein level
- 13 January 2010
- journal article
- research article
- Published by Cold Spring Harbor Laboratory in RNA
- Vol. 16 (3), 489-494
- https://doi.org/10.1261/rna.1701210
Abstract
MRNA profiling is routinely used to identify microRNA targets, however, this high-throughput technology is not suitable for identifying targets regulated only at protein level. Here, we have developed and validated a novel methodology based on computational analysis of promoter sequences combined with mRNA microarray experiments to reveal transcription factors that are direct microRNA targets at the protein level. Using this approach we identified Smad3, a key transcription factor in the TGFβ signaling pathway, as a direct miR-140 target. We showed that miR-140 suppressed the TGFβ pathway through repression of Smad3 and that TGFβ suppressed the accumulation of miR-140 forming a double negative feedback loop. Our findings establish a valid strategy for the discovery of microRNA targets regulated only at protein level, and we propose that additional targets could be identified by re-analysis of existing microarray datasets.Keywords
This publication has 28 references indexed in Scilit:
- Emerging Role of miR-106b-25/miR-17-92 Clusters in the Control of Transforming Growth Factor β SignalingCancer Research, 2008
- A crucial role for GW182 and the DCP1:DCP2 decapping complex in miRNA-mediated gene silencingRNA, 2005
- MicroRNA Expression in Zebrafish Embryonic DevelopmentScience, 2005
- Serum response factor regulates a muscle-specific microRNA that targets Hand2 during cardiogenesisNature, 2005
- Principles of MicroRNA–Target RecognitionPLoS Biology, 2005
- Microarray analysis shows that some microRNAs downregulate large numbers of target mRNAsNature, 2005
- MicroRNAs: Genomics, Biogenesis, Mechanism, and FunctionCell, 2004
- The Basic Helix-Loop-Helix Factor, HAND2, Functions as a Transcriptional Activator by Binding to E-boxes as a HeterodimerPublished by Elsevier BV ,2002
- TGF-β/Smad3 Signals Repress Chondrocyte Hypertrophic Differentiation and Are Required for Maintaining Articular CartilageThe Journal of cell biology, 2001
- Direct binding of Smad3 and Smad4 to critical TGFbeta -inducible elements in the promoter of human plasminogen activator inhibitor-type 1geneThe EMBO Journal, 1998