title: Mir-20a regulates in vitro mineralization and BMP signaling pathway by targeting BMP-2 transcript in fish.
abstract: MicroRNAs (miRNAs) are important regulators of vertebrate development but their role during skeletogenesis remains unknown. In this regard, we investigated the mineralogenic activity of miR-20a, a miRNA associated with osteogenesis, in fish bone-derived cells. Expression of miR-20a was up-regulated during differentiation and its overexpression inhibited mineralization, suggesting a role in fish tissue calcification. In this regard, a conserved miR-20a binding site was identified in bone morphogenetic protein 2 (BMP-2) 3'UTR and its functionality was evidenced through luciferase assays, and further confirmed by western-blot and qPCR. Type II BMP receptor (BMPR2) is also targeted by miR-20a in mammalian systems and evidence was collected for the presence of a binding site in fish sequences. We propose that miR-20a is a regulator of BMP pathway through specific action on BMP-2 and possibly BMPR2. Overexpression of miR-20a was also shown to up-regulate matrix Gla protein (MGP) transcript, a physiological inhibitor of calcification previously found to form a complex with BMP-2. We propose that MGP may play a role in the anti-mineralogenic effect promoted by miR-20a by decreasing availability of BMP-2. This study gives new insights into miRNA-mediated regulation of BMP-2, and sheds light into the potential role of miR-20a as a regulator of skeletogenesis.
authors.affiliation: Centre of Marine Sciences (CCMAR/CIMAR-LA), University of Algarve, Faro, Portugal. Electronic address: dtiago@ualg.pt.
authors.forename: Daniel M
authors.initials: DM
authors.lastname: Tiago
authors.affiliation: Centre of Marine Sciences (CCMAR/CIMAR-LA), University of Algarve, Faro, Portugal; PhD Program in Biomedical Sciences, University of Algarve, Faro, Portugal.
authors.forename: Cátia L
authors.initials: CL
authors.lastname: Marques
authors.affiliation: Centre of Marine Sciences (CCMAR/CIMAR-LA), University of Algarve, Faro, Portugal; PhD Program in Biomedical Sciences, University of Algarve, Faro, Portugal.
authors.forename: Vânia P
authors.initials: VP
authors.lastname: Roberto
authors.affiliation: Centre of Marine Sciences (CCMAR/CIMAR-LA), University of Algarve, Faro, Portugal; Department of Biomedical Sciences and Medicine, University of Algarve, Faro, Portugal.
authors.forename: M Leonor
authors.initials: ML
authors.lastname: Cancela
authors.affiliation: Centre of Marine Sciences (CCMAR/CIMAR-LA), University of Algarve, Faro, Portugal.
authors.forename: Vincent
authors.initials: V
authors.lastname: Laizé
mesh_terms: D000818:Animals; D001665:Binding Sites; D055396:Bone Morphogenetic Protein 2 / Q000235:genetics* / Q000378:metabolism*; D002113:Calcification, Physiologic / Q000235:genetics*; D002460:Cell Line; D017124:Conserved Sequence; D019143:Evolution, Molecular; D005109:Extracellular Matrix / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D035683:MicroRNAs / Q000235:genetics*; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D021541:Sea Bream; D015398:Signal Transduction / Q000235:genetics*; D015854:Up-Regulation; D000519:alpha-Galactosidase / Q000378:metabolism
chemical_list: D055396:Bone Morphogenetic Protein 2; D035683:MicroRNAs; D012333:RNA, Messenger; C576330:GLA protein, human; D000519:alpha-Galactosidase
keywords: BMP-2; MGP-BMP-2 interaction; Mineralizing fish bone-derived cells; Post-transcriptional regulation; miR-20a