Identification and characterization of novel ER-based hsp90 gene in the red flour beetle, Tribolium castaneum

Abstract

Heat-shock protein 90 (HSP90) is a highly conserved molecular chaperone found in all species except for Archaea, which is required not only for stress tolerance but also for normal development. Recently, it was reported that HSP83, one member of the cytosolic HSP90 family, contributes to oogenesis and responds to heat resistance in Tribolium castaneum. Here, a novel ER-based HSP90 gene, Tchsp90, has been identified in T. castaneum. Phylogenetic analysis showed that hsp90s and hsp83s evolved separately from a common ancestor but that hsp90s originated earlier. Quantitative real-time polymerase chain reaction illustrated that Tchsp90 is expressed in all developmental stages and is highly expressed at early pupa and late adult stages. Tchsp90 was upregulated in response to heat stress but not to cold stress. Laval RNAi revealed that Tchsp90 is important for larval/ pupal development. Meanwhile, parental RNAi indicated that it completely inhibited female fecundity and partially inhibited male fertility once Tchsp90 was knocked down and that it will further shorten the lifespan of T. castaneum. These results suggest that Tchsp90 is essential for development, lifespan, and reproduction in T. castaneum in addition to its response to heat stress.

Characterization and functional analysis of hsp21.8b: An orthologous small heat shock protein gene in Tribolium castaneum

“…As T. castaneum is basically living in the warm cereal storage place and without diapause, Tchsp21.8b was in response to heat but exhibited insensitiveness to cold stress. This phenotype was also existed in ER‐based Tchsp90 and Tchsp18.3 , both of which were only upregulated by heat stress (Xie et al., ; Zhang, Gu et al., ), indicating that the behaviour would make a similar induction of other hsps in T. castaneum .…”

“…This result suggested that the orthologous shsp also could be in response of the oxidative stress caused by UV exposure, similar with hsp70 and hsp90 . Likewise, the orthologous shsp showed analogous expression profiles with hsp70 and hsp90 under starvation stress (Shim, Kim, & Lee, ; Zhang, Gu et al., ). As both of hsp70 level and hsp90 level decreased greatly when the insect larvae were starved for days, Tchsp21.8b was also downregulated by starvation stress for 3 days, with a decreased degree of 20.8%–41.8% compared to the control (Figure b).…”

“…PCR product was used as template for dsRNA synthesis with the TranscriptAid™ T7 High Yield Transcription Kit (Fermentas, Vilnius, Lithuania). The purification, solution and quality checking of dsRNA were performed as described previously (Zhang, Gu et al., ).…”

“…During the evolution history, a series of proteins arose to protect cells against stress conditions, such as heat, cold, oxidation, pathogen stress and so on (Richter, Haslbeck, & Buchner, ; Zhang, Gu et al., ). A prominent group of these stress proteins are the molecular chaperones, which named as heat shock proteins (HSPs) and classified into several families according to their molecular mass and evolutionary history (Haslbeck & Vierling, ; Kim, Kim, & Kim, ).…”

Real‐time cell analysis and heat shock protein gene expression in the TcA Tribolium castaneum cell line in response to environmental stress conditions

“…Additionally, they have a significant role in the protection and maintenance of many fundamental cellular functions (Nadeau et al ., ). There are several studies on Hsps characterization and expression in response to stress conditions in T. castaneum (Xu et al ., ; Holmstrup et al ., ; Schinko et al ., ; Zhang et al ., ). Sang et al .…”