Identification of proteasome subunit beta type 2 associated with deltamethrin detoxification in Drosophila Kc cells by cDNA microarray analysis and bioassay analyses

Gene. 2016 May 10;582(1):85-93. doi: 10.1016/j.gene.2016.01.054. Epub 2016 Feb 2.

Abstract

Insecticide deltamethrin resistance has presented a difficult obstacle for pest control and the resistance development is complex and associated with many genes. To better understand the possible molecular mechanisms involved in DM stress, in this study, cDNA microarray analysis was employed. 448 differentially expressed genes with at least a 2-fold expression difference were identified in Drosophila cells after DM exposure. Moreover, some genes were confirmed with qPCR, which yielded results consistent with the microarray analysis. Three members of the ubiquitin-proteasome system were significantly elevated in DM-stressed cells, suggesting that the ubiquitin-proteasome pathway may play an important role in DM detoxification. The proteasome beta2 subunit (Prosbeta2) is a member of 20S proteasome subunit family, which forms the proteolytic core of 26S proteasome. Whether Prosbeta2 participates in DM detoxification requires further study. RNAi and heterologous expression were conducted to investigate the contribution of Prosbeta2 in DM detoxification. The results revealed Prosbeta2 knockdown significantly reduce the level of DM detoxification in RNAi-treated cells after 48 h. Overexpression of Prosbeta2 increased cellular viability. These detoxification results represent the first evidence that Prosbeta2 plays a role in the detoxification of DM, which may provide new idea and target for studying the molecular mechanisms of insect resistance.

Keywords: Deltamethrin resistance; Detoxification; Heterologous expression; Overexpression; Prosbeta2; RNAi; cDNA microarray.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Drosophila / drug effects
  • Drosophila / genetics
  • Drosophila / metabolism
  • Drosophila Proteins / biosynthesis*
  • Drosophila Proteins / genetics
  • Endopeptidases / biosynthesis*
  • Endopeptidases / genetics
  • Gene Expression Regulation / drug effects
  • Gene Knockdown Techniques
  • Insecticide Resistance / drug effects*
  • Insecticide Resistance / genetics
  • Nitriles / toxicity
  • Oligonucleotide Array Sequence Analysis / methods
  • Proteasome Endopeptidase Complex / chemistry
  • Proteasome Endopeptidase Complex / genetics*
  • Pyrethrins / toxicity

Substances

  • Drosophila Proteins
  • Nitriles
  • Pyrethrins
  • decamethrin
  • Endopeptidases
  • Prosbeta2 protein, Drosophila
  • Proteasome Endopeptidase Complex