Insecticide resistance mediated by an exon skipping event

Many genes increase coding capacity by alternate exon usage. The gene encoding the insect nicotinic acetylcholine receptor (nAChR) α6 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 s...

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Main Authors:	Berger, Madeleine, Puinean, Alin Mirel, Randall, Emma, Zimmer, Christoph T., Silva, Wellington M., Bielza, Pablo, Field, Linda M., Hughes, David, Mellor, Ian, Hassani‐Pak, Keywan, Siqueira, Herbert A. A., Williamson, Martin S., Bass, Chris
Format:	Online
Language:	English
Published:	John Wiley and Sons Inc. 2016
Online Access:	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5111602/

id	pubmed-5111602
recordtype	oai_dc
spelling	pubmed-51116022016-11-16 Insecticide resistance mediated by an exon skipping event Berger, Madeleine Puinean, Alin Mirel Randall, Emma Zimmer, Christoph T. Silva, Wellington M. Bielza, Pablo Field, Linda M. Hughes, David Mellor, Ian Hassani‐Pak, Keywan Siqueira, Herbert A. A. Williamson, Martin S. Bass, Chris ORIGINAL ARTICLES Many genes increase coding capacity by alternate exon usage. The gene encoding the insect nicotinic acetylcholine receptor (nAChR) α6 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 Taα6 resulting in a novel form of insecticide resistance mediated by exon skipping. Sequencing of the α6 subunit cDNA from spinosad selected and unselected strains of T. absoluta revealed all Taα6 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 α7, a model of insect α6, 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 α6 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. John Wiley and Sons Inc. 2016-11-02 2016-11 /pmc/articles/PMC5111602/ /pubmed/27748560 http://dx.doi.org/10.1111/mec.13882 Text en © 2016 The Authors. Molecular Ecology Published by John Wiley & Sons Ltd. This is an open access article under the terms of the Creative Commons Attribution (http://creativecommons.org/licenses/by/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
repository_type	Open Access Journal
institution_category	Foreign Institution
institution	US National Center for Biotechnology Information
building	NCBI PubMed
collection	Online Access
language	English
format	Online
author	Berger, Madeleine Puinean, Alin Mirel Randall, Emma Zimmer, Christoph T. Silva, Wellington M. Bielza, Pablo Field, Linda M. Hughes, David Mellor, Ian Hassani‐Pak, Keywan Siqueira, Herbert A. A. Williamson, Martin S. Bass, Chris
spellingShingle	Berger, Madeleine Puinean, Alin Mirel Randall, Emma Zimmer, Christoph T. Silva, Wellington M. Bielza, Pablo Field, Linda M. Hughes, David Mellor, Ian Hassani‐Pak, Keywan Siqueira, Herbert A. A. Williamson, Martin S. Bass, Chris Insecticide resistance mediated by an exon skipping event
author_facet	Berger, Madeleine Puinean, Alin Mirel Randall, Emma Zimmer, Christoph T. Silva, Wellington M. Bielza, Pablo Field, Linda M. Hughes, David Mellor, Ian Hassani‐Pak, Keywan Siqueira, Herbert A. A. Williamson, Martin S. Bass, Chris
author_sort	Berger, Madeleine
title	Insecticide resistance mediated by an exon skipping event
title_short	Insecticide resistance mediated by an exon skipping event
title_full	Insecticide resistance mediated by an exon skipping event
title_fullStr	Insecticide resistance mediated by an exon skipping event
title_full_unstemmed	Insecticide resistance mediated by an exon skipping event
title_sort	insecticide resistance mediated by an exon skipping event
description	Many genes increase coding capacity by alternate exon usage. The gene encoding the insect nicotinic acetylcholine receptor (nAChR) α6 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 Taα6 resulting in a novel form of insecticide resistance mediated by exon skipping. Sequencing of the α6 subunit cDNA from spinosad selected and unselected strains of T. absoluta revealed all Taα6 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 α7, a model of insect α6, 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 α6 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.
publisher	John Wiley and Sons Inc.
publishDate	2016
url	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5111602/
_version_	1613729278164205568

Insecticide resistance mediated by an exon skipping event

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