Control of Dopa decarboxylase gene expression by the Broad-Complex during metamorphosis in Drosophila.
The induction of the Dopa decarboxylase gene (Ddc) in the epidermis of Drosophila at pupariation is a receptor-mediated response to the steroid molting hormone, ecdysone. Activity is also dependent on the Broad-Complex (BR-C), an early ecdysone response gene that functions during metamorphosis. BR-C encodes a family of zinc-finger protein isoforms, BR-C(Z1-Z4). Genetic experiments have shown that the Z2 isoform is required for epidermal Ddc to reach maximum expression at pupariation. In this paper, we report that BR-C regulates Ddc expression at two different developmental stages through two different cis-acting regions. At pupariation, BR-C acts synergistically with the ecdysone receptor to up-regulate Ddc. DNase I foot printing has identified four binding sites of the predominant Z2 isoform within a distal regulatory element that is required for maximal Ddc activity. The sites share a conserved core sequence with a set of BR-C sites that had been mapped previously to within the first Ddc intron. Using variously deleted Ddc genomic regions to drive reporter gene expression in transgenic organisms, we show that the intronic binding sites are required for Ddc expression at eclosion. At both pupariation and eclosion, BR-C releases Ddc from an active silencing mechanism, operating through two distinct cis-acting regions of the Ddc genomic domain at these stages. Transgenes, bearing a Ddc fragment from which one of the cis-acting silencers has been deleted, exhibit beta-galactosidase reporter activity in the epidermal cells prior to the appearance of endogenous DDC. Our finding that BR-C is required for Ddc activation at eclosion is the first evidence to suggest that this important regulator of the early metamorphic events, also regulates target gene expression at the end of metamorphosis.
Chen
Li
L
Department of Biological Sciences, University of Alberta, Edmonton, AB T6G 2E9, Canada.
O'Keefe
Sandra L
SL
Hodgetts
Ross B
RB
D017398:Alternative Splicing; D000818:Animals; D001665:Binding Sites; D002467:Cell Nucleus / Q000378:metabolism; D003433:Crosses, Genetic; D003850:Deoxyribonuclease I / Q000378:metabolism; D004296:Dopa Decarboxylase / Q000096:biosynthesis*; D004330:Drosophila; D029721:Drosophila Proteins / Q000096:biosynthesis / Q000737:chemistry*; D004440:Ecdysone / Q000494:pharmacology; D000078404:Epidermal Cells; D018507:Gene Expression Regulation, Developmental; D015971:Gene Expression Regulation, Enzymologic; D020868:Gene Silencing; D017930:Genes, Reporter; D007438:Introns; D008675:Metamorphosis, Biological; D008957:Models, Genetic; D009154:Mutation; D010957:Plasmids / Q000378:metabolism; D020033:Protein Isoforms; D014157:Transcription Factors / Q000096:biosynthesis / Q000737:chemistry*
C102117:Br protein, Drosophila; D029721:Drosophila Proteins; D020033:Protein Isoforms; D014157:Transcription Factors; D004440:Ecdysone; D003850:Deoxyribonuclease I; D004296:Dopa Decarboxylase