RNA interference of NADPH-cytochrome P450 reductase results in reduced insecticide resistance in the bed bug, Cimex lectularius

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

doi:10.1371/journal.pone.0031037

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RNA interference of NADPH-cytochrome P450 reductase results in reduced insecticide resistance in the bed bug, Cimex lectularius

Journal article

Open access

RNA interference of NADPH-cytochrome P450 reductase results in reduced insecticide resistance in the bed bug, Cimex lectularius

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

PloS one, Vol.7(2), pp.e31037-e31037

2012

DOI: https://doi.org/10.1371/journal.pone.0031037

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https://hdl.handle.net/2376/108792

PMCID: PMC3274526

PMID: 22347424

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Abstract

NADPH-Ferrihemoprotein Reductase - genetics

Nitriles - pharmacology

Computational Biology

Insecticide Resistance - genetics

NADPH-Ferrihemoprotein Reductase - chemistry

Insecticides

Phylogeny

Pyrethrins - pharmacology

Bedbugs - drug effects

Tissue Distribution

Animals

RNA Interference

Insecticide Resistance - drug effects

Binding Sites

Bedbugs - enzymology

DNA, Complementary

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.

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Title: RNA interference of NADPH-cytochrome P450 reductase results in reduced insecticide resistance in the bed bug, Cimex lectularius
Creators: Fang Zhu - Department of Entomology, College of Agriculture, University of Kentucky, Lexington, Kentucky, United States of America
Sarah Sams
Tim Moural
Kenneth F Haynes
Michael F Potter
Subba R Palli
Publication Details: PloS one, Vol.7(2), pp.e31037-e31037
Publisher: United States
Identifiers: 99900547038301842
Language: English
Resource Type: Journal article