A spinosyn-sensitive Drosophila melanogaster nicotinic acetylcholine receptor identified through chemically induced target site resistance, resistance gene identification, and heterologous expression.
Strains of Drosophila melanogaster with resistance to the insecticides spinosyn A, spinosad, and spinetoram were produced by chemical mutagenesis. These spinosyn-resistant strains were not cross-resistant to other insecticides. The two strains that were initially characterized were subsequently found to have mutations in the gene encoding the nicotinic acetylcholine receptor (nAChR) subunit Dalpha6. Subsequently, additional spinosyn-resistant alleles were generated by chemical mutagenesis and were also found to have mutations in the gene encoding Dalpha6, providing convincing evidence that Dalpha6 is a target site for the spinosyns in D. melanogaster. Although a spinosyn-sensitive receptor could not be generated in Xenopus laevis oocytes simply by expressing Dalpha6 alone, co-expression of Dalpha6 with an additional nAChR subunit, Dalpha5, and the chaperone protein ric-3 resulted in an acetylcholine- and spinosyn-sensitive receptor with the pharmacological properties anticipated for a native nAChR.
Watson
Gerald B
GB
Dow AgroSciences LLC, Discovery Research, 9330 Zionsville Road, Indianapolis, IN 46268, USA. gbwatson@dow.com
Chouinard
Scott W
SW
Cook
Kevin R
KR
Geng
Chaoxian
C
Gifford
Jim M
JM
Gustafson
Gary D
GD
Hasler
James M
JM
Larrinua
Ignacio M
IM
Letherer
Ted J
TJ
Mitchell
Jon C
JC
Pak
William L
WL
Salgado
Vincent L
VL
Sparks
Thomas C
TC
Stilwell
Geoff E
GE
D000818:Animals; D018833:Chaperonins / Q000235:genetics / Q000378:metabolism; D029721:Drosophila Proteins / Q000235:genetics* / Q000378:metabolism*; D004331:Drosophila melanogaster; D004338:Drug Combinations; D004351:Drug Resistance / Q000187:drug effects / Q000235:genetics*; D015870:Gene Expression; D007306:Insecticides / Q000494:pharmacology*; D018942:Macrolides / Q000494:pharmacology*; D009154:Mutation; D009865:Oocytes / Q000166:cytology / Q000378:metabolism; D011978:Receptors, Nicotinic / Q000235:genetics* / Q000378:metabolism*; D014982:Xenopus laevis
D029721:Drosophila Proteins; D004338:Drug Combinations; D007306:Insecticides; D018942:Macrolides; D011978:Receptors, Nicotinic; C488972:nAChRalpha6 protein, Drosophila; C114883:spinosyn A; D018833:Chaperonins; C415329:spinosad; C548262:spinetoram