In Drosophila the steroid hormone ecdysone triggers a genetic regulatory hierarchy in which ecdysone combines with a receptor protein to form a complex that induces the transcription of a small class of "early" genes, which encode transcription factors that regulate other genes. We previously reported that one of the early genes, E75, encodes members of the steroid receptor superfamily. Using an E75 hybridization probe, we have identified two additional Drosophila genes that encode members of this superfamily. One of these is the ecdysone receptor gene, EcR, as previously reported. In this work, we examine the sequence, genomic organization, and developmental expression of the other gene, DHR3, which, like E75, encodes one of a growing number of "orphan" receptors for which ligands have not yet been identified. The structure of the DHR3 protein is strikingly similar to that of the MHR3 protein (e.g., 97% amino acid identity for the DNA binding domains), another orphan receptor encoded by an ecdysone-inducible early gene of another insect, Manduca sexta. The temporal developmental profile for DHR3 expression closely parallels that for the ecdysone titer and for the ecdysone-inducible E75 and E74 Drosophila early genes. The structural similarity to a Manduca early gene and the expression similarities to Drosophila early genes suggest that the DHR3 gene may also belong to the early gene class.