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The ecdysone receptor (EcR) and ultraspiracle (USP) belong to a large class of nuclear hormone receptors that are activated by insect molting hormones. In this study, we describe the isolation of the Bombyx mori EcR and USP genes and their localization on the chromosomal map of the silkworm. The predicted amino acid sequences of the EcR and USP proteins are highly conserved between insects. Genomic analysis of the B. mori EcR and USP genes showed that both genes contain two transcriptional units. The relative levels of USP transcripts were higher than those of EcR, and the transcripts showed a different distribution within the different organs examined. The temporal and spatial expression patterns of the EcR gene were consistent with the phenotypic characteristics of B. mori larvae. EcR transcripts accumulated during larval molting and the molt cycle. EcR mRNA was shown to be localized in the midgut of the silkworm. Moreover, the EcR protein was present in the cytoplasm of midgut cells, and after a short pulse of ecdysteroid treatment the EcR protein was translocated to the nucleus. These results suggest that the EcR protein function in the regulation of gene expression during ecdysis. We examined the activity of EcR in conjunction with USP in a heterologous system. Recombinant EcR and USP proteins showed the ability to activate transcription in a dose-dependent manner by binding to endogenous cis-acting elements in Drosophila melanogaster genomic DNA. When the specific binding site was mutated, the EcR protein, in combination with the USP protein, failed to activate transcription. These results indicate that EcR and USP are involved in the ecdysone response of D. melanogaster and that these two proteins can act both alone and in combination to activate transcription in the same D. melanogaster genomic DNA region.Q:
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