Transcriptional regulation of BMP4 synexpression in transgenic Xenopus.

Synexpression groups are genetic modules composed of genes that share both a complex expression pattern and the biological process in which they function. Here we investigate the regulation of BMP4 synexpression by studying the enhancers of bambi, smad7 and vent2 in Xenopus. We find that a BMP4 synexpression promoter module is compact and (i) requires direct BMP responsiveness through Smad and Smad-cofactor binding motifs, (ii) may contain an evolutionary conserved BMP-responsive element, bre7 (TGGCGCC), that is crucial for expression of bambi and smad7 and is highly prognostic for novel BMP-responsive enhancers (BREs); and (iii) requires a narrow window of BMP inducibility, because minor enhancement or reduction of BMP responsiveness abolishes synexpression. Furthermore, we used a bioinformatic model to predict in silico 13 novel BREs, and tested five of them that were found in the id1-4 genes. The results highlight that in vivo analysis is required to reveal the physiological, spatio-temporal regulation of BMP-responsive genes.