title: EXPRESSION AND EFFECTS OF MUTANT Bombyx mori ACETYLCHOLINESTRASE1 IN BmN CELLS.
abstract: The main mechanism of toxicity of organophosphate (OP) and carbamate (CB) insecticides is their irreversible binding and inhibition of acetylcholinestrase (AChE), encoded by ace1 (acetylcholinestrase gene 1), leading to eventual death of insects. Mutations in AChE may significantly reduce insects susceptibility to these pesticides. Bombyx mori is an important beneficial insect, and no OP- or CB-resistant strains have been generated. In this study, wild-type ace1 (wace1) and mutant ace1 (mace1) were introduced into BmN cells, confirmed by screening and identification. The expression of wace1 and mace1 in the cells was confirmed by Western blot and their expression levels were about 21-fold higher than the endogenous ace1 level. The activities of AChE in wace1 and mace1 transgenic cells were 10.6 and 20.2% higher compared to control cells, respectively. mace1 transgenic cells had higher remaining activity than wace1 transgenic cells under the treatment of physostigmine (a reversible cholinesterase inhibitor) and phoxim (an OP acaricide). The results showed that ace1 transgene can significantly improve ace1 expression, and ace1 mutation at a specific site can reduce the sensitivity to AChE inhibitors. Our study provides a new direction for the exploration of the relationship between AChE mutations and drug resistance.
authors.affiliation: School of Basic Medicine and Biological Sciences, Soochow University, Suzhou, Jiangsu, P. R. China.
authors.forename: Bin-Bin
authors.initials: BB
authors.lastname: Wang
authors.affiliation: School of Basic Medicine and Biological Sciences, Soochow University, Suzhou, Jiangsu, P. R. China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Xie
authors.affiliation: School of Basic Medicine and Biological Sciences, Soochow University, Suzhou, Jiangsu, P. R. China.
authors.forename: Fan-Chi
authors.initials: FC
authors.lastname: Li
authors.affiliation: School of Basic Medicine and Biological Sciences, Soochow University, Suzhou, Jiangsu, P. R. China.
authors.forename: Min
authors.initials: M
authors.lastname: Ni
authors.affiliation: School of Basic Medicine and Biological Sciences, Soochow University, Suzhou, Jiangsu, P. R. China.
authors.forename: Kai-Zun
authors.initials: KZ
authors.lastname: Xu
authors.affiliation: School of Basic Medicine and Biological Sciences, Soochow University, Suzhou, Jiangsu, P. R. China.
authors.forename: Jiang-Hai
authors.initials: JH
authors.lastname: Tian
authors.affiliation: School of Basic Medicine and Biological Sciences, Soochow University, Suzhou, Jiangsu, P. R. China.
authors.forename: Jing-Sheng
authors.initials: JS
authors.lastname: Hu
authors.affiliation: School of Basic Medicine and Biological Sciences, Soochow University, Suzhou, Jiangsu, P. R. China.
authors.forename: Bin
authors.initials: B
authors.lastname: Xue
authors.affiliation: School of Basic Medicine and Biological Sciences, Soochow University, Suzhou, Jiangsu, P. R. China.|National Engineering Laboratory for Modern Silk, Soochow University, Suzhou, Jiangsu, P. R. China.
authors.forename: Wei-De
authors.initials: WD
authors.lastname: Shen
authors.affiliation: School of Basic Medicine and Biological Sciences, Soochow University, Suzhou, Jiangsu, P. R. China. lib@suda.edu.cn.|National Engineering Laboratory for Modern Silk, Soochow University, Suzhou, Jiangsu, P. R. China. lib@suda.edu.cn.
authors.forename: Bing
authors.initials: B
authors.lastname: Li
mesh_terms: D000110:Acetylcholinesterase / Q000235:genetics* / Q000378:metabolism*; D000818:Animals; D012831:Bombyx / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics*; D002460:Cell Line; D002800:Cholinesterase Inhibitors / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects; D019476:Insect Proteins / Q000235:genetics* / Q000378:metabolism*; D007306:Insecticides / Q000494:pharmacology; D009154:Mutation; D009946:Organothiophosphorus Compounds / Q000494:pharmacology; D010830:Physostigmine / Q000494:pharmacology; D014162:Transfection
chemical_list: D002800:Cholinesterase Inhibitors; D019476:Insect Proteins; D007306:Insecticides; D009946:Organothiophosphorus Compounds; C003135:phoxim; D010830:Physostigmine; D000110:Acetylcholinesterase
keywords: Bombyx mori; acetylcholinestrase; enzyme activity; mutation; transgenetic