Spatial regulation of Corazonin neuropeptide expression requires multiple cis-acting elements in Drosophila melanogaster

Abstract

Although most invertebrate neuropeptide-encoding genes display distinct expression patterns in the central nervous system (CNS), the molecular mechanisms underlying spatial regulation of the neuropeptide genes are largely unknown. Expression of the neuropeptide Corazonin (Crz) is limited to only 24 neurons in the larval CNS of Drosophila melanogaster, and these neurons have been categorized into three groups, namely, DL, DM, and vCrz. To identify cis-regulatory elements that control transcription of Crz in each neuronal group, reporter gene expression patterns driven by various 5' flanking sequences of Crz were analyzed to assess their promoter activities in the CNS. We show that the 504-bp 5' upstream sequence is the shortest promoter directing reporter activities in all Crz neurons. Further dissection of this sequence revealed two important regions responsible for group specificity: -504::-419 for DM expression and -380::-241 for DL and vCrz expression. The latter region is further subdivided into three sites (proximal, center, and distal), in which any combinations of the two are sufficient for DL expression, whereas both proximal and distal sites are required for vCrz expression. Interestingly, the TATA box does not play a role in Crz transcription in most neurons. We also show that a 434-bp 5' upstream sequence of the D. virilis Crz gene, when introduced into the D. melanogaster genome, drives reporter expression in the DL and vCrz neurons, suggesting that regulatory mechanisms for Crz expression in at least two such neuronal groups are conserved between the two species.