Insecticide resistance mediated by an exon skipping event.

A - Papers appearing in refereed journals

Berger, M., Puinean, A. M., Randall, E., Zimmer, C. T., Silva, W. M., Bielza, P., Field, L. M., Hughes, D. J., Mellor, I., Hassani-Pak, K., Siqueira, H. A. A., Williamson, M. S. and Bass, C. 2016. Insecticide resistance mediated by an exon skipping event. Molecular Ecology. 25 (22), pp. 5692-5704.

AuthorsBerger, M., Puinean, A. M., Randall, E., Zimmer, C. T., Silva, W. M., Bielza, P., Field, L. M., Hughes, D. J., Mellor, I., Hassani-Pak, K., Siqueira, H. A. A., Williamson, M. S. and Bass, C.
Abstract

Many genes increase coding capacity by alternate exon usage. The gene encoding the insect nicotinic acetylcholine receptor (nAChR) a6 subunit, target of the bio-insecticide spinosad, is one example of this and expands protein diversity via alternative splicing of mutually exclusive exons. Here, we show that spinosad resistance in the tomato leaf miner, Tuta absoluta is associated with aberrant regulation of splicing of Ta alpha 6 resulting in a novel form of insecticide resistance mediated by exon skipping. Sequencing of the alpha 6 subunit cDNA from spinosad selected and unselected strains of T. absoluta revealed all Ta alpha 6 transcripts of the selected strain were devoid of exon 3, with comparison of genomic DNA and mRNA revealing this is a result of exon skipping. Exon skipping cosegregated with spinosad resistance in survival bioassays, and functional characterization of this alteration using modified human nAChR alpha 7, a model of insect alpha 6, demonstrated that exon 3 is essential for receptor function and hence spinosad sensitivity. DNA and RNA sequencing analyses suggested that exon skipping did not result from genetic alterations in intronic or exonic cis-regulatory elements, but rather was associated with a single epigenetic modification downstream of exon 3a, and quantitative changes in the expression of trans-acting proteins that have known roles in the regulation of alternative splicing. Our results demonstrate that the intrinsic capacity of the alpha 6 gene to generate transcript diversity via alternative splicing can be readily exploited during the evolution of resistance and identifies exon skipping as a molecular alteration conferring insecticide resistance.

KeywordsBiochemistry & Molecular Biology; Ecology; Evolutionary Biology
Year of Publication2016
JournalMolecular Ecology
Journal citation25 (22), pp. 5692-5704
Digital Object Identifier (DOI)doi:10.1111/mec.13882
PubMed ID27748560
Open accessPublished as ‘gold’ (paid) open access
FunderPeople Programme (Marie Curie Actions) of the European Union under REA grant [PIRSES-GA-2012 - 318246]
Biotechnology and Biological Sciences Research Council
Funder project or codeSustainability
BBSRC Quota Studentship: Investigating the molecular mechanisms of insecticide resistance in the tomato leafminer Tuta absoluta
A genomic approach to understanding insecticide resistance in crop pests
1096240
Publisher's version
Copyright license
CC BY
File Access Level
Open
Output statusPublished
Publication dates
Online17 Oct 2016
Publication process dates
Accepted05 Oct 2016
PublisherWiley
ISSN0962-1083

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