Identification of transducer molecules for BMP signaling

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1. Identification of signal transducers binding to the cytoplasmic region of BMP type I receptor (BMPR-I). To clarify BMP receptor mediated signaling, the cytoplasmic interactors cDNAs of BMPR-I,which includes serine-threonine kinase, were isolated from human placenta cDNA library with the ose of a yeast two-hydrid system. The isolated 4 clones were termed to BRAM1-4 (BMP receptor associated molecule 1-4). We confirmed that BRAM2 interacted to catalytic domain of BMPR-I in yeast and mammalian cells. Moreover, BRAM2 also interacted to TGF-beta type I receptor.2. Identification of BMP-inducing molecules cDNA.To identify target genes for BMP signaling, we performed Differential display method using the mRNA prepared from MC3T3-E1 cells with or without BMP-stimulation, and Dlx-5 gene, which contains a homeotic box, were isolated. We confirmed that Dlx-5 gene was expressed by BMP-stimulation. The stable cell line which overexpressed Dlx-5, seems to lead the bone differentiation.