Submitted to: Archives of Insect Biochemistry and Physiology
Publication Type: Peer reviewed journal
Publication Acceptance Date: 10/28/2008
Publication Date: 2/1/2009
Citation: An, S., Wang, S., Stanley, D.W., Song, Q. 2009. Identification of a Novel Bursicon-regulated Transcriptional Regulator, md133790, in House Fly Musca domestica. Archives of Insect Biochemistry and Physiology. 70:106-121. Interpretive Summary: Use of classical insecticides has introduced severe problems in agricultural and environmental sustainability. Two of most pressing problems are the ability of pest insects to resist classical insecticides and the negative influence of insecticides on ecosystems. One approach to reduce the environmental insecticide load is based on the concept of disrupting insect hormone systems to cripple insect development. This idea grew from discovery that chemicals that naturally occur in a few plant species block insect development. Although this modern approach to insect control is feasible, direct application of this approach is problematic. The concept of disrupting insect development is limited by lack of detailed knowledge of insect hormone systems. To help solve this problem, we are investigating how one of the insect hormones influences insect development. In this paper we report on identification of a specific hormone effect. We show that the hormone regulates expression of a specific gene required for development. This new research will be directly useful to scientists who are working to improve the usefulness of disrupting insect development. The ensuing improved methods will benefit a wide range of agricultural producers, and people who rely on agricultural products, by supporting the long-term sustainability of agriculture.
Technical Abstract: Bursicon is a neuropeptide that regulates cuticle sclerotization (hardening and tanning) and wing expansion in insects via a G-protein coupled receptor. The peptide consists of alpha and beta subunits. In the present study, we cloned bursicon alpha and beta genes in the house fly Musca domestica using 3’ and 5’ RACE and expressed the recombinant bursicon (rbursicon) heterodimer in mammalian 293 cells and insect Highfive**TM cells. The rbursicon displayed a strong bursicon activity in the neck-ligated house fly assay. Using rbursicon, we identified and cloned a novel bursicon-regulated gene in M. domestica encoding a transcriptional regulator homologous to ataxin-7-like3 in human, CG13379 in Drosophila and sgf11in yeast Saccharomyces cerevisiae. We named the gene md13379. Both ataxin-7-like3 and sgf11 are a novel subunit of the SAGA (Spt-Ada-Gcn5-Acetyltransferase) complex that is involved in regulation of gene transcription. Real-time PCR analysis of temporal response profile revealed that the level of md13379 transcript was up-regulated by 6.6 fold 1h after rbursicon injection, which correlates well with the cuticle sclerotization process observed in the rbursicon-injected flies. The composite data suggest that md13379 plays a role in regulating the expression of bursicon-regulated genes involved in the cuticle sclerotization process.