SmTR2/4, a Schistosoma mansoni homologue of TR2/TR4 orphan nuclear receptor.

cDNA clones encoding a Schistosoma mansoni homologue of the TR2/TR4 group of nuclear receptors, SmTR2/4, were identified by screening an adult female worm cDNA library. SmTR2/4 is a 1,943 amino acid protein, the largest member of the TR2/TR4 group of nuclear receptors and also the largest nuclear receptor reported to date. SmTR2/4 retains a typical domain organisation of nuclear receptors exhibiting 69-77% sequence identity in the DNA binding domain and 16-22% sequence identity in the ligand binding domain compared with its orthologues. SmTR2/4 contains a large A/B domain and hinge region. SmTR2/4 also contains a 100 amino acid F domain, which is absent from its orthologues. SmTR2/4 mRNA is expressed in every stage of the S. mansoni life cycle, exhibiting an elevated expression level in cercariae. Western blot analysis identified two forms of SmTR2/4 protein in adult worms. Our in vitro DNA binding assay showed that SmTR2/4 binds to the DR-3 consensus hormone response element, suggesting a functional conservation among the TR2/TR4 group members in terms of DNA binding specificity. A yeast-based transactivation assay demonstrated that the A/B domain, F domain and N-terminal part of the hinge region in SmTR2/4, when tethered to a GAL4 DNA binding domain, exhibited an autonomous transcription activation function.