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Steroid hormones are crucial for many biological events in multicellular organisms. In insects, the principal steroid hormones are ecdysteroids, which play essential roles in regulating molting and metamorphosis. During larval and pupal development, ecdysteroids are synthesized in the prothoracic gland (PG) from dietary cholesterol via a series of hydroxylation and oxidation steps. The expression of all but one of the known ecdysteroid biosynthetic enzymes is restricted to the PG, but the transcriptional regulatory networks responsible for generating such exquisite tissue-specific regulation is only beginning to be elucidated. Here, we report identification and characterization of the C 2 H 2 -type zinc finger transcription factor Ouija board (Ouib) necessary for ecdysteroid production in the PG in the fruit fly  Drosophila melanogaster . Expression of  ouib  is predominantly limited to the PG, and genetic null mutants of  ouib  result in larval developmental arrest that can be rescued by administrating an active ecdysteroid. Interestingly,  ouib  mutant animals exhibit a strong reduction in the expression of one ecdysteroid biosynthetic enzyme,  spookier . Using a cell culture-based luciferase reporter assay, Ouib protein stimulates transcription of  spok  by binding to a specific ~15 bp response element in the  spok  PG enhancer element. Most remarkable, the developmental arrest phenotype of  ouib  mutants is rescued by over-expression of a functionally-equivalent paralog of  spookier . These observations imply that the main biological function of Ouib is to specifically regulate  spookier  transcription during  Drosophila  development.

The Drosophila Zinc Finger Transcription Factor Ouija Board Controls Ecdysteroid Biosynthesis through Specific Regulation of spookier