The same amino acid substitution in orthologous esterases confers organophosphate resistance on the house fly and a blowfly.
Organophosphate (OP) insecticide resistance in certain strains of Musca domestica is associated with reduction in the carboxylesterase activity of a particular esterase isozyme. This has been attributed to a 'mutant ali-esterase hypothesis', which invokes a structural mutation to an ali-esterase resulting in the loss of its carboxylesterase activity but acquisition of OP hydrolase activity. It has been shown that the mutation in Lucilia cuprina is a Gly137-->Asp substitution in the active site of an esterase encoded by the Lc alpha E7 gene (Newcomb, R.D., Campbell, P.M., Ollis, D.L., Cheah, E., Russell, R.J., Oakeshott, J.G., 1997. A single amino acid substitution converts a carboxylesterase to an organophosphate hydrolase and confers insecticide resistance on a blowfly. Proc. Natl. Acad. Sci. USA 94, 7464-7468). We now report the cloning and characterisation of the orthologous M. domestica Md alpha E7 gene, including the sequencing of cDNAs from the OP resistant Rutgers and OP susceptible sbo and WHO strains. The Md alpha E7 gene has the same intron structure as Lc alpha E7 and encodes a protein with 76% amino acid identity to Lc alpha E7. Comparisons between susceptible and resistance alleles show resistance in M. domestica is associated with the same Gly137-->Asp mutation as in L. cuprina. Bacterial expression of the Rutgers allele shows its product has OP hydrolase activity. The data indicate identical catalytic mechanisms have evolved in orthologous Md alpha E7 and Lc alpha E7 molecules to endow diazinon-type resistance on the two species of higher Diptera.
Claudianos
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Division of Entomology, Commonwealth Scientific and Industrial Research Organisation, Canberra, Australia. charlesc@ento.csiro.au
Russell
R J
RJ
Oakeshott
J G
JG
D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000818:Animals; D001483:Base Sequence; D018076:DNA, Complementary; D004175:Diptera / Q000201:enzymology*; D004950:Esterases / Q000145:classification / Q000235:genetics* / Q000378:metabolism; D006793:Houseflies / Q000201:enzymology*; D006868:Hydrolysis; D007305:Insecticide Resistance; D007306:Insecticides; D008969:Molecular Sequence Data; D009943:Organophosphorus Compounds; D017386:Sequence Homology, Amino Acid
D018076:DNA, Complementary; D007306:Insecticides; D009943:Organophosphorus Compounds; D004950:Esterases