Comparative RNA-sequencing analysis of ER-based HSP90 functions and signal pathways in Tribolium castaneum

Abstract

Tribolium castaneum, the red flour beetle, is a major agriculture pest that damages stored grains and cereal products. Heat-shock protein 90 of T. castaneum (Tchsp90) has been reported to play pivotal roles in heat stress response, development, reproduction, and life span. However, the signaling pathway of Tchsp90 remains unclear. Thus, the global transcriptome profiles between RNA interference (RNAi)-treated insects (ds-Tchsp90) and control insects of T. castaneum were investigated and compared by RNA sequencing. In all, we obtained 14,145,451 sequence reads, which assembled into 13,243 genes. Among these genes, 461 differentially expressed genes (DEGs) were identified between the ds-Tchsp90 and control samples. These DEGs were classified into 44 gene ontology (GO) functional groups, including the cellular process, the response to stimulus, the immune system process, the development process, and reproduction. Interestingly, knocking down the expression of Tchsp90 suppressed both the DNA replication and cell division signaling pathways, which most likely modulated the effects of Tchsp90 on development, reproduction, and life span. Moreover, the DEGs encoding AnnexinB9, frizzled-4, sno, Fem1B, TSL, and CSW might be related to the regulation of the development and reproduction of ds-Tchsp90 insects. The DEGs including TLR6, PGRP2, defensin1, and defensin2 were involved in heat stress and immune response simultaneously, which suggested that cross talk might exist between immunity and stress response. Additionally, RNAi of Tchsp90 altered large-scale serine protease (sp) gene expression patterns and amplified the SP signaling pathway to regulate the development and reproduction as well as the stress response and innate immunity in T. castaneum. All these results shed new light onto the regulatory mechanism of Tchsp90 involved in insect physiology and could further facilitate research into appropriate and sustainable pest control management.