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Published online 1 June 2004. doi:10.1083/jcb.200311057
The Rockefeller University Press, 0021-9525 $8.00
JCB, Volume 165, Number 5, 631-640
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Article

 Ecdysone receptor directly binds the promoter of the Drosophila caspase dronc, regulating its expression in specific tissues

Dimitrios Cakouros1, Tasman J. Daish1,2, and Sharad Kumar1,2

1 Hanson Institute, Institute of Medical and Veterinary Science, Adelaide, SA 5000, Australia
2 Department of Medicine, Adelaide University, Adelaide, SA 5005, Australia

Address correspondence to S. Kumar, Haematology, Hanson Institute, Institute of Medical and Veterinary Science, Frome Rd., Adelaide, SA 5000, Australia. Tel.: 61-8-82223738. Fax: 61-8-8222-3139. email: sharad.kumar{at}imvs.sa.gov.au

The steroid hormone ecdysone regulates moulting, cell death, and differentiation during insect development. Ecdysone mediates its biological effects by either direct activation of gene transcription after binding to its receptor EcR–Usp or via hierarchical transcriptional regulation of several primary transcription factors. In turn, these transcription factors regulate the expression of several downstream genes responsible for specific biological outcomes. DRONC, the Drosophila initiator caspase, is transcriptionally regulated by ecdysone during development. We demonstrate here that the dronc promoter directly binds EcR–Usp. We further show that mutation of the EcR–Usp binding element (EcRBE) reduces transcription of a reporter and abolishes transactivation by an EcR isoform. We demonstrate that EcRBE is required for temporal regulation of dronc expression in response to ecdysone in specific tissues. We also uncover the participation of a putative repressor whose function appears to be coupled with EcR–Usp. These results indicate that direct binding of EcR–Usp is crucial for controlling the timing of dronc expression in specific tissues.

Key Words: nuclear hormone receptors; gene transcription; steroid hormones; apoptosis; transcriptional regulation

Abbreviations used in this paper: EcRBE, EcR–Usp binding element; EMSA, electrophoretic mobility shift analysis; PCD, programmed cell death; TSA, Trichostatin A.


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