title: Cloning and characterization of prophenoloxidase A3 (proPOA3) from Culex pipiens pallens.
abstract: The prophenoloxidase subunit A3 (proPOA3) gene was cloned from Culex pipiens pallens, which had an open reading frame of 2061 bp encoding a putative 686 amino acid protein. The deduced amino acid sequence shares 98% with proPOA3 from Culex quinquefasciatus. ProPOA3 is expressed at all developmental stages of C. pipiens pallens. Significant negative correlation was observed between proPOA3 expression and deltamethrin resistance in resistant C. pipiens pallens. Furthermore, proPOA3 expression levels were significantly lower in deltamethrin-resistant mosquitoes than in susceptible mosquitoes collected at four locations in Eastern China. However, we did not find any substantial change in proPOA3 expression in field-collected resistant Anopheles mosquitoes. Moreover, overexpressing proPOA3 in C6/36 cells led to more sensitivity to deltamethrin treatment. In laboratory and field-collected resistant C. pipiens pallens, a valine to isoleucine mutation (769G>A) and two synonymous mutations (1116G>C and 1116G>A) were identified in proPOA3. In addition, the mutation frequency of 769G>A and 1116G>C increased gradually, which corresponded with raised deltamethrin resistance levels. Taken together, our study provides the first evidence that proPOA3 may play a role in the regulation of deltamethrin-resistance in C. pipiens pallens.
authors.affiliation: Department of Pathogen Biology, Nanjing Medical University, Nanjing, Jiangsu 210029, PR China.
authors.forename: Dan
authors.initials: D
authors.lastname: Zhou
authors.forename: Shunhua
authors.initials: S
authors.lastname: Hao
authors.forename: Yan
authors.initials: Y
authors.lastname: Sun
authors.forename: Lin
authors.initials: L
authors.lastname: Chen
authors.forename: Chunrong
authors.initials: C
authors.lastname: Xiong
authors.forename: Lei
authors.initials: L
authors.lastname: Ma
authors.forename: Donghui
authors.initials: D
authors.lastname: Zhang
authors.forename: Shanchao
authors.initials: S
authors.lastname: Hong
authors.forename: Linna
authors.initials: L
authors.lastname: Shi
authors.forename: Maoqing
authors.initials: M
authors.lastname: Gong
authors.forename: Huayun
authors.initials: H
authors.lastname: Zhou
authors.forename: Xinyou
authors.initials: X
authors.lastname: Yu
authors.forename: Bo
authors.initials: B
authors.lastname: Shen
authors.forename: Changliang
authors.initials: C
authors.lastname: Zhu
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D004156:Catechol Oxidase / Q000096:biosynthesis / Q000235:genetics*; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D003001:Cloning, Molecular; D003465:Culex / Q000201:enzymology* / Q000254:growth & development; D004351:Drug Resistance; D004792:Enzyme Precursors / Q000096:biosynthesis / Q000235:genetics*; D015870:Gene Expression; D018507:Gene Expression Regulation, Developmental; D005787:Gene Frequency; D019476:Insect Proteins / Q000096:biosynthesis / Q000235:genetics*; D007306:Insecticides / Q000494:pharmacology; D007928:Lethal Dose 50; D008969:Molecular Sequence Data; D009570:Nitriles / Q000494:pharmacology; D017354:Point Mutation; D011722:Pyrethrins / Q000494:pharmacology; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D017422:Sequence Analysis, DNA; D013045:Species Specificity
chemical_list: D004792:Enzyme Precursors; D019476:Insect Proteins; D007306:Insecticides; D009570:Nitriles; D011722:Pyrethrins; D011994:Recombinant Proteins; C017180:decamethrin; C038451:pro-phenoloxidase; D004156:Catechol Oxidase