Molecular characterization and RNA interference analysis of the DEAD-box gene family in Locusta migratoria.

Abstract DEAD-box (DDX) genes encode a group of RNA helicases that are highly conserved and ubiquitously expressed from prokaryotes to eukaryotes, and appear to participate in almost every aspect of RNA metabolism. Studies have been extensively done in yeast and human, in insect, beyond the flies, however, the information of these genes is limited. Here, we therefore identified and characterized 32 DDX genes from Locusta migratoria (L. migratoria), a crop pest. Overview of the gene structure and domain composition showed that the gene size varies significantly from one to fifteen exons, and the encoded proteins contain the conserved helicase core with various extensions at their amino and carboxyl termini. Phylogenetic trees informed that these locust DDX family members have orthologs in all insect species examined and can be classified into 30 subfamilies, all of them found counterparts in human, and most in yeast as well. Quantitative real-time PCR revealed that these genes are expressed in all stages and tissues examined, overall with higher expression level at second and third-instar nymphs and in the reproductive organs. RNA interference (RNAi) analyses showed that seven genes cause lethal phenotype when silenced, of which five lead to defective midgut and gastric caecum, indicating that these genes are essential for the survival and maintenance of normal digestive organs of locust. These data provide a foundation for further functional analysis of these DDX genes in locust.