title: Resistance-associated point mutations of organophosphate insensitive acetylcholinesterase, in the olive fruit fly Bactrocera oleae.
abstract: A 2.2-kb full length cDNA containing an ORF encoding a putative acetylcholinesterase (AChE) precursor of 673 amino acid residues was obtained by a combined degenerate PCR and RACE strategy from an organophosphate-susceptible Bactrocera oleae strain. A comparison of cDNA sequences of individual insects from susceptible and resistant strains, coupled with an enzyme inhibition assay with omethoate, indicated a novel glycine-serine substitution (G488S), at an amino acid residue which is highly conserved across species (G396 of Torpedocalifornica AChE), as a likely cause of AChE insensitivity. This mutation was also associated with a 35-40% reduction in AChE catalytic efficiency. The I199V substitution, which confers low levels of resistance in Drosophila, was also present in B. oleae (I214V) and in combination with G488S produced up to a 16-fold decrease in insecticide sensitivity. This is the first agricultural pest where resistance has been associated with an alteration in AChE, which arises from point mutations located within the active site gorge of the enzyme.
authors.affiliation: Liverpool School of Tropical Medicine, Pembroke Place, Liverpool L3 5QA, UK. vontas@imbb.forth.gr
authors.forename: J G
authors.initials: JG
authors.lastname: Vontas
authors.forename: M J
authors.initials: MJ
authors.lastname: Hejazi
authors.forename: N J
authors.initials: NJ
authors.lastname: Hawkes
authors.forename: N
authors.initials: N
authors.lastname: Cosmidis
authors.forename: M
authors.initials: M
authors.lastname: Loukas
authors.forename: R W
authors.initials: RW
authors.lastname: Janes
authors.forename: J
authors.initials: J
authors.lastname: Hemingway
mesh_terms: D000110:Acetylcholinesterase / Q000235:genetics* / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D002800:Cholinesterase Inhibitors; D004117:Dimethoate / Q000031:analogs & derivatives*; D004175:Diptera / Q000201:enzymology* / Q000235:genetics; D007305:Insecticide Resistance / Q000235:genetics; D007306:Insecticides; D008958:Models, Molecular; D008969:Molecular Sequence Data; D017354:Point Mutation; D011487:Protein Conformation; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid
chemical_list: D002800:Cholinesterase Inhibitors; D007306:Insecticides; C002302:dimethoxon; D000110:Acetylcholinesterase; D004117:Dimethoate