Transcriptional regulation of gilthead seabream bone morphogenetic protein (BMP) 2 gene by bone- and cartilage-related transcription factors

Abstract

Bone morphogenetic protein (BMP) 2 belongs to the transforming growth factor β (TGFβ) superfamily of cytokines and growth factors. While it plays important roles in embryo morphogenesis and organogenesis, BMP2 is also critical to bone and cartilage formation. Protein structure and function have been remarkably conserved throughout evolution and BMP2 transcription has been proposed to be tightly regulated, although few data is available. In this work we report the cloning and functional analysis of gilthead seabream BMP2 promoter. As in other vertebrates, seabream BMP2 gene has a 5′ non-coding exon, a feature already present in DPP gene, the fruit fly ortholog of vertebrate BMP2 gene, and maintained throughout evolution. In silico analysis of seabream BMP2 promoter revealed several binding sites for bone and cartilage related transcription factors (TFs) and their functionality was evaluated using promoter-luciferase constructions and TF-expressing vectors. Runt-related transcription factor 3 (RUNX3) was shown to negatively regulate BMP2 transcription and combination with the core binding factor β (CBFβ) further reduced transcriptional activity of the promoter. Although to a lesser extent, myocyte enhancer factor 2C (MEF2C) had also a negative effect on the regulation of BMP2 gene transcription, when associated with SRY (sex determining region Y)-box 9 (SOX9b). Finally, v-ets avian erythroblastosis virus E26 oncogene homolog 1 (ETS1) was able to slightly enhance BMP2 transcription. Data reported here provides new insights toward the better understanding of the transcriptional regulation of BMP2 gene in a bone and cartilage context.

Evidences for a New Role of miR-214 in Chondrogenesis

“…In fish, the role of Ets1 in skeleton formation remains to be explored. Nevertheless, in gilthead seabream this TF was shown to regulate the expression of two important regulators of cartilage formation: bmp2 and cartilage-specific s100 32 , 33 . Altogether, these results indicate that ETS1 could be important to control vertebrate skeletogenesis through a putative regulation of Dnm3os/ miR-214 expression.…”

RGS19 upregulates Nm23-H1/2 metastasis suppressors by transcriptional activation via the cAMP/PKA/CREB pathway

“…The presence of transcription factor binding sites in the putative promoter regions of NME1/2 was predicted using the MatInspector version 7.1 [ 51 ] at www.genomatix.de . Sites with scores (core similarity) below 0.75 were not considered [ 52 ]. The NME1 promoter sequence was from GenBank: L35301, L34822; the NME2 promoter sequence (2000 bp upstream of 5’ flanking sequence) was from Ensemble (NME2 ENSG00000243678).…”

Mef2 and the skeletal muscle differentiation program

“…Chick Mef2c is expressed early and broadly in mesoderm, suggesting it may affect development of various tissues [25]. Over-expression of zebrafish Mef2cs in heterologous systems may influence osteo-or chondrogenesis through Sox9/BMP [65]. After initial myogenesis, zebrafish mef2ca mRNA becomes localised to fibre ends, whereas Xenopus Mef2c has been suggested to function in early tendon cells [29,66].…”