title: Functional characterization of NADPH-cytochrome P450 reductase from Bactrocera dorsalis: Possible involvement in susceptibility to malathion.
abstract: NADPH cytochrome P450 reductase (CPR) is essential for cytochrome P450 catalysis, which is important in the detoxification and activation of xenobiotics. In this study, two transcripts of Bactrocera dorsalis CPR (BdCPR) were cloned, and the deduced amino-acid sequence had an N-terminus membrane anchor for BdCPR-X1 and three conserved binding domains (FMN, FAD, and NADP), as well as an FAD binding motif and catalytic residues for both BdCPR-X1 and BdCPR-X2. BdCPR-X1 was detected to have the high expression levels in adults and in Malpighian tubules, fat bodies, and midguts of adults, but BdCPR-X2 expressed lowly in B. dorsalis. The levels of BdCPRs were similar in malathion-resistant strain compared to susceptible strain. However, injecting adults with double-stranded RNA against BdCPR significantly reduced the transcript levels of the mRNA, and knockdown of BdCPR increased adult susceptibility to malathion. Expressing complete BdCPR-X1 cDNA in Sf9 cells resulted in high activity determined by cytochrome c reduction and these cells had higher viability after exposure to malathion than control. The results suggest that BdCPR could affect the susceptibility of B. dorsalis to malathion and eukaryotic expression of BdCPR would lay a solid foundation for further investigation of P450 in B. dorsalis.
authors.affiliation: Key Laboratory of Entomology and Pest Control Engineering, College of Plant Protection, Southwest University, Chongqing 400716, P. R. China.
authors.forename: Yong
authors.initials: Y
authors.lastname: Huang
authors.affiliation: Key Laboratory of Entomology and Pest Control Engineering, College of Plant Protection, Southwest University, Chongqing 400716, P. R. China.
authors.forename: Xue-Ping
authors.initials: XP
authors.lastname: Lu
authors.affiliation: Key Laboratory of Entomology and Pest Control Engineering, College of Plant Protection, Southwest University, Chongqing 400716, P. R. China.
authors.forename: Luo-Luo
authors.initials: LL
authors.lastname: Wang
authors.affiliation: Key Laboratory of Entomology and Pest Control Engineering, College of Plant Protection, Southwest University, Chongqing 400716, P. R. China.
authors.forename: Dong
authors.initials: D
authors.lastname: Wei
authors.affiliation: Key Laboratory of Entomology and Pest Control Engineering, College of Plant Protection, Southwest University, Chongqing 400716, P. R. China.
authors.forename: Zi-Jiao
authors.initials: ZJ
authors.lastname: Feng
authors.affiliation: Key Laboratory of Entomology and Pest Control Engineering, College of Plant Protection, Southwest University, Chongqing 400716, P. R. China.
authors.forename: Qi
authors.initials: Q
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Entomology and Pest Control Engineering, College of Plant Protection, Southwest University, Chongqing 400716, P. R. China.
authors.forename: Lin-Fan
authors.initials: LF
authors.lastname: Xiao
authors.affiliation: Key Laboratory of Entomology and Pest Control Engineering, College of Plant Protection, Southwest University, Chongqing 400716, P. R. China.
authors.forename: Wei
authors.initials: W
authors.lastname: Dou
authors.affiliation: Key Laboratory of Entomology and Pest Control Engineering, College of Plant Protection, Southwest University, Chongqing 400716, P. R. China.
authors.forename: Jin-Jun
authors.initials: JJ
authors.lastname: Wang
mesh_terms: D000818:Animals; D003001:Cloning, Molecular; D045304:Cytochromes c / Q000378:metabolism; D005260:Female; D007305:Insecticide Resistance / Q000235:genetics*; D007306:Insecticides / Q000633:toxicity*; D008294:Malathion / Q000633:toxicity*; D008297:Male; D009251:NADPH-Ferrihemoprotein Reductase / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D010802:Phylogeny; D020033:Protein Isoforms / Q000145:classification / Q000235:genetics / Q000378:metabolism; D034622:RNA Interference; D012330:RNA, Double-Stranded / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D017422:Sequence Analysis, DNA; D061987:Sf9 Cells; D018411:Spodoptera; D033621:Tephritidae / Q000187:drug effects* / Q000201:enzymology*; D059467:Transcriptome
chemical_list: D007306:Insecticides; D020033:Protein Isoforms; D012330:RNA, Double-Stranded; D012333:RNA, Messenger; D045304:Cytochromes c; D009251:NADPH-Ferrihemoprotein Reductase; D008294:Malathion