Cloning and expression of a tauropine dehydrogenase from the marine sponge Suberites domuncula

Abstract

The cDNA sequence coding for tauropine dehydrogenase (TaDH) [belonging to the family of opine dehydrogenases] has been determined. Using the demosponge Suberites domuncula, we describe for the first time the tauropine dehydrogenase gene (of length 2,992 kb) from a eukaryote, consisting of two introns flanked by three exons. Moreover, two allelic variants have been identified, which are present in the different specimens either in a homozygotic or in a heterozygotic way; the data suggest an intermediary type of heritance. Phylogenetic analyses indicate that S. domuncula TaDH is only distantly related to the opine dehydrogenases from marine invertebrates; rather it comprises high sequence similarity to bacterial ornithine cyclodeaminases (OCD). In addition, expression studies revealed that the steady-state level of TaDH dropped drastically in animals, which had been exposed to elevated aeration. Antibodies raised against the recombinant sponge TaDH were used to demonstrate that S. domuncula expresses high levels of this enzyme in almost all cells. If tissue samples were kept under additional aeration no immuno-signals could be identified. A strong accumulation of the enzyme was seen around the bacteria, existing in bacteriocytes, indicating that under aerobic conditions the bacteria might produce taurine. These data suggest involvement of the sponge TaDH in the final step of the glycolytic pathway, more specifically, in regeneration of NAD(+) under anaerobic conditions. Furthermore, potential mutual influences between bacteria and sponge are discussed, claiming a horizontal gene transfer of the gene from a bacterium to the sponge. © 2008 Springer-Verlag.