CncC regulated genes code for proteins involved in all three phases of insecticide detoxification in Tribolium Castaneum

In invertebrates, a heterodimer of xenobiotic transcription factors, cap n collar C isoform (CncC) and muscle aponeurosis fibromatosis (Maf) mediate cellular defense. In insects, these proteins regulate expression of genes involved in insecticide detoxification. In the current study, we performed sequencing of RNA isolated from Tribolium castaneum pyrethriod resistant strain (QTC279) beetles injected with CncC or green fluroscence protein (GFP, control) dsRNA. Differential expression analysis of RNA sequences identified 662 genes that showed a decrease and 91 genes that showed an increase in expression (at a p value 1.5) in CncC knockdown insects when compared to their expression in control insects. We focused on the downregulated genes and selected a subset of 27 genes (21 with a predicted function in xenobiotic detoxification and six randomly picked) and verified their differential expression using qRT-PCR. RNAi and insecticide bioassays were employed to study the function of six of these genes coding for CYP4G7, CYP4G14, GST-1 and four ABC transporters, ABCA-UB, ABCA-A1 and ABCA-A1L and ABCA-9B involved in all three phases of insecticide detoxification. These data suggest that CncC is a major regulator of genes coding for proteins involved in detoxification of insecticides.In invertebrates, a heterodimer of xenobiotic transcription factors, cap n collar C isoform (CncC) and muscle aponeurosis fibromatosis (Maf) mediate cellular defense. In insects, these proteins regulate expression of genes involved in insecticide detoxification. In the current study, we performed sequencing of RNA isolated from Tribolium castaneum pyrethriod resistant strain (QTC279) beetles injected with CncC or green fluroscence protein (GFP, control) dsRNA. Differential expression analysis of RNA sequences identified 662 genes that showed a decrease and 91 genes that showed an increase in expression (at a p value 1.5) in CncC knockdown insects when compared to their expression in control insects. We focused on the downregulated genes and selected a subset of 27 genes (21 with a predicted function in xenobiotic detoxification and six randomly picked) and verified their differential expression using qRT-PCR. RNAi and insecticide bioassays were employed to study the function of six of these genes coding for CYP4G7, CYP4G14, GST-1 and four ABC transporters, ABCA-UB, ABCA-A1 and ABCA-A1L and ABCA-9B involved in all three phases of insecticide detoxification. These data suggest that CncC is a major regulator of genes coding for proteins involved in detoxification of insecticides.

在无脊椎动物中，异源二聚体形式的异生物转录因子——帽颈C同型（CncC）与肌腱纤维瘤（Maf）——介导细胞防御机制。在昆虫中，这些蛋白质调节参与杀虫剂解毒的基因表达。在本研究中，我们对从拟南芥虫 Tribolium castaneum 的抗拟除虫菊酯菌株（QTC279）中分离出的RNA进行了测序，这些虫子分别被注射了CncC或绿色荧光蛋白（GFP，对照组）的双链RNA。RNA序列的差异表达分析识别出662个基因表达量降低，91个基因表达量增加（p值1.5），与对照昆虫相比，在CncC敲低昆虫中表现出差异。我们聚焦于下调基因，并从其中选取了27个基因子集（其中21个具有预测的异生物解毒功能，其余6个随机选取）并使用实时定量PCR验证了它们差异表达。通过RNA干扰和杀虫剂生物测定，研究了其中6个基因的功能，这些基因编码的蛋白质包括CYP4G7、CYP4G14、GST-1和四个ABC转运蛋白，即ABCA-UB、ABCA-A1、ABCA-A1L和ABCA-9B，它们参与杀虫剂解毒的三个阶段。这些数据表明，CncC是调控参与杀虫剂解毒的蛋白质编码基因的主要调节因子。