Broad-complex Z3 contributes to the ecdysone-mediated transcriptional regulation of the vitellogenin gene in Bombus lantschouensis.

During reproduction, vitellogenin (Vg), as an egg yolk precursor, is critical in sexually mature females of oviparous species including some insects. The transcription of Vg is usually mediated by hormones such as juvenile hormone (JH), ecdysteroids and some neuropeptides. In this study, the structure of the Vg gene from the bumblebee Bombus lantschouensis, (BlVg) was determined by sequencing and assembly. BlVg was found to be expressed at higher levels in reproductive queens than in virgins by quantitative real-time PCR analysis. Tissue-specific expression analysis showed that BlVg was expressed at the highest levels in the fat bodies of both virgin and reproductive queens. Prediction of the BlVg promoter revealed the presence of ecdysteroid-responsive cis-regulatory elements (CREs) containing one Broad-Complex zinc-finger isoform 3 (BR-C Z3), and one ecdysone-induced protein 74A (E74A). In addition, luciferase reporter expression, driven by the 5' -regulatory region of the BlVg gene, from -1517 bp to +895 bp downstream of the start codon, was induced by treatment with 20-hydroxyecdysone (20-E). Moreover, the luciferase activity of the BlVg promoter was elevated by only BlBrC-Z3 when Sf9 cells were cotransfected with four BlBrC isoforms respectively. BlVg promoter-mediated luciferase activation was significantly reduced when the putative BrC-Z3 CRE in the promoter was mutated. In summary, this report describes the first study of vitellogenin gene regulation at the transcriptional level in bumblebees and demonstrates that the ecdysone-induced transcription of the BlVg gene is mediated by the binding of BlBrC-Z3 to the BrC-Z3 CRE in the BlVg promoter in bumblebees.