Drosophila nicotinic acetylcholine receptor subunits and their native interactions with insecticidal peptide toxins.

Autor:	Korona D; Department of Genetics, University of Cambridge, Downing Street, Cambridge, United Kingdom., Dirnberger B; Department of Genetics, University of Cambridge, Downing Street, Cambridge, United Kingdom.; Cambridge Centre for Proteomics, Department of Biochemistry, University of Cambridge, Cambridge, United Kingdom.; Syngenta, Jealott's Hill International Research Centre, Bracknell, United Kingdom., Giachello CNG; Syngenta, Jealott's Hill International Research Centre, Bracknell, United Kingdom., Queiroz RML; Cambridge Centre for Proteomics, Department of Biochemistry, University of Cambridge, Cambridge, United Kingdom., Popovic R; MRC Toxicology Unit, Gleeson Building, University of Cambridge, Tennis Court Road, Cambridge, United Kingdom., Müller KH; Cambridge Advanced Imaging Centre, Department of Physiology, Development and Neuroscience/Anatomy Building, University of Cambridge, Cambridge, United Kingdom., Minde DP; Cambridge Centre for Proteomics, Department of Biochemistry, University of Cambridge, Cambridge, United Kingdom., Deery MJ; Cambridge Centre for Proteomics, Department of Biochemistry, University of Cambridge, Cambridge, United Kingdom., Johnson G; Department of Genetics, University of Cambridge, Downing Street, Cambridge, United Kingdom., Firth LC; Syngenta, Jealott's Hill International Research Centre, Bracknell, United Kingdom., Earley FG; Syngenta, Jealott's Hill International Research Centre, Bracknell, United Kingdom., Russell S; Department of Genetics, University of Cambridge, Downing Street, Cambridge, United Kingdom., Lilley KS; Cambridge Centre for Proteomics, Department of Biochemistry, University of Cambridge, Tennis Court Road, Cambridge, United Kingdom.
Jazyk:	angličtina
Zdroj:	ELife [Elife] 2022 May 16; Vol. 11. Date of Electronic Publication: 2022 May 16.
DOI:	10.7554/eLife.74322
Abstrakt:	Drosophila nicotinic acetylcholine receptors (nAChRs) are ligand-gated ion channels that represent a target for insecticides. Peptide neurotoxins are known to block nAChRs by binding to their target subunits, however, a better understanding of this mechanism is needed for effective insecticide design. To facilitate the analysis of nAChRs we used a CRISPR/Cas9 strategy to generate null alleles for all ten nAChR subunit genes in a common genetic background. We studied interactions of nAChR subunits with peptide neurotoxins by larval injections and styrene maleic acid lipid particles (SMALPs) pull-down assays. For the null alleles, we determined the effects of α-Bungarotoxin (α-Btx) and ω-Hexatoxin-Hv1a (Hv1a) administration, identifying potential receptor subunits implicated in the binding of these toxins. We employed pull-down assays to confirm α-Btx interactions with the Drosophila α5 (D α 5), Dα6, D α 7 subunits. Finally, we report the localisation of fluorescent tagged endogenous Dα6 during Drosophila CNS development. Taken together, this study elucidates native Drosophila nAChR subunit interactions with insecticidal peptide toxins and provides a resource for the in vivo analysis of insect nAChRs. Competing Interests: DK, CG, RQ, RP, KM, DM, MD, GJ, FE, SR, KL No competing interests declared, BD, LF is affiliated with Syngenta. The author has no other competing interests to declare (© 2022, Korona et al.)
Databáze:	MEDLINE
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