Journal Article

RNA Interference of NADPH-Cytochrome P450 Reductase Results in Reduced Insecticide Resistance in the Bed Bug, Cimex lectularius

Moural, Tim, Haynes, Kenneth F., Sams, Sarah, Zhu, Fang, Palli, Subba R., Potter, Michael F.

2012

Overview

NADPH-cytochrome P450 reductase (CPR) plays a central role in cytochrome P450 action. The genes coding for P450s are not yet fully identified in the bed bug, Cimex lectularius. Hence, we decided to clone cDNA and knockdown the expression of the gene coding for CPR which is suggested to be required for the function of all P450s to determine whether or not P450s are involved in resistance of bed bugs to insecticides. The full length Cimex lectularius CPR (ClCPR) cDNA was isolated from a deltamethrin resistant bed bug population (CIN-1) using a combined PCR strategy. Bioinformatics and in silico modeling were employed to identify three conserved binding domains (FMN, FAD, NADP), a FAD binding motif, and the catalytic residues. The critical amino acids involved in FMN, FAD, NADP binding and their putative functions were also analyzed. No signal peptide but a membrane anchor domain with 21 amino acids which facilitates the localization of ClCPR on the endoplasmic reticulum was identified in ClCPR protein. Phylogenetic analysis showed that ClCPR is closer to the CPR from the body louse, Pediculus humanus corporis than to the CPRs from the other insect species studied. The ClCPR gene was ubiquitously expressed in all tissues tested but showed an increase in expression as immature stages develop into adults. We exploited the traumatic insemination mechanism of bed bugs to inject dsRNA and successfully knockdown the expression of the gene coding for ClCPR. Suppression of the ClCPR expression increased susceptibility to deltamethrin in resistant populations but not in the susceptible population of bed bugs. These data suggest that P450-mediated metabolic detoxification may serve as one of the resistance mechanisms in bed bugs.

• Publisher: Public Library of Science,Public Library of Science (PLoS)
• Subject: Adults / Agriculture / Agrochemicals / Amino acids / Animals / Bed bug (Cimex lectularius) / Bedbugs - drug effects / Bedbugs - enzymology / Binding / Binding Sites / Bioinformatics / Biology / Catalysis / Cimex lectularius / Cladistic analysis / Cloning / Computational Biology / Cytochrome / Cytochrome P450 / Cytochromes P450 / Data processing / Deltamethrin / Detoxification / DNA, Complementary / Double-stranded RNA / Endoplasmic reticulum / Enzymes / Flavin mononucleotide / Flavin-adenine dinucleotide / Gene expression / Genes / Insecticide resistance / Insecticide Resistance - drug effects / Insecticide Resistance - genetics / Insecticides / Insects / Laboratories / Localization / Medicine / Metabolism / Mutation / NADP / NADPH-cytochrome P450 reductase / NADPH-ferrihemoprotein reductase / NADPH-Ferrihemoprotein Reductase - chemistry / NADPH-Ferrihemoprotein Reductase - genetics / Nitriles - pharmacology / Pediculus humanus / Pesticide resistance / Phylogeny / Polymerase chain reaction / Population / Pyrethrins - pharmacology / Reductases / Ribonucleic acid / RNA / RNA Interference / RNA-mediated interference / Signal peptides / Studies / Tissue Distribution / Tissues