Bone Morphogenetic Protein-4-induced Activation of Xretpos is Mediated by Smads and Olf-1/EBF Associated Zinc Finger (OAZ)

We have previously isolated Xretpos, a novel family of long terminal repeat (LTR)-retrotransposons in Xenopus laevis, whose transcript is restricted to ventro-posterior-specific regions and induced by bone morphogenetic protein-4 (BMP-4) signaling. To explore the molecular mechanism of the transcriptional regulation, we identified and characterized Xretpos promoter regions consisting of LTRs and a 5'-untranslated region. We demonstrated that this promoter region contains all the necessary regulatory elements for the spatial and temporal expression of XRETPOS: Sequence analysis of the Xretpos promoter revealed multiple Smad-binding elements and Olf-1/EBF-associated zinc finger (OAZ) binding sites similar to BMP-4 response element, which were identified and proved to be required for BMP-4 induction in the Xvent2 promoter. We further demonstrated that Smads and OAZ proteins bind to their response elements in the promoter and these bindings are essential for the BMP-4-induced activation of the Xretpos promoter. Furthermore, we showed that the endogenous expression of Xretpos protein indeed occurred and was temporally regulated and BMP-4-inducible during the early Xenopus development. Finally, overexpression and partial loss-of-function study revealed that Xretpos has a posterio-ventralizing activity. Together, our results place Xretpos downstream of BMP-4 and provide evidence for the conserved mechanism of transcriptional regulation of the BMP-4 target genes.