Expression pattern of the Brachyury and Tbx2 homologues from the sponge Suberites domuncula

T-box transcription factors are a large family of transcriptional regulators involved in many aspects of embryonic development. In a previous report, we described the isolation and genomic characterization of two T-box genes from the siliceous sponge Suberites domuncula: a Brachyury homologue, Sd-Bra, and a Tbx2 homologue, Sd-Tbx2. Elucidation of the genomic structure of Sd-Bra allowed us to demonstrate the existence of two different isoforms, resulting from alternative splicing. Moreover, we demonstrated that the shorter isoform exists in two different glycosylation states.

In the present study, we demonstrate a differential subcellular localization of the three Sd-Bra isoforms, suggesting that its differential nuclear import could be an important mechanism for its functional regulation. Furthermore, we demonstrate that Sd-Tbx2 exists only in one isoform, which is mainly localized in the nucleus. The pattern of expression of Sd-Bra and Sd-Tbx2 genes is analysed in sponge tissue, in gemmules and in cultured cells.

These results suggest a conserved role for Sd-Bra in the control of morphogenetic movements through the regulation of cell-adhesion properties and the involvement of Sd-Tbx2 in the determination of cell identity in the early stages of differentiation, reminiscent of the function of Tbx2-3-4-5 in vertebrates during limb specification. Also, the fact that a Brachyury and a Tbx2 homologue exist in S. domuncula suggests that the first divergence from the ancestral Brachyury-like gene might be a Tbx2-like gene and not a Tbrain-like gene as had been previously suggested.