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Polymorphisms in GSTE2 is associated with temephos resistance in Aedes aegypti

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Helvecio, Elisama, Romão, Tatiany Patrícia, de Carvalho Leandro, Danilo, Ferreira de Oliveira, Iêda, Holanda D. Cavalcanti, Emanuel, Reimer, Lisa ORCID: https://orcid.org/0000-0002-9711-4981, de Paiva Cavalcanti, Milena, Silva de Oliveira, Ana Patrícia, Guedes Paiva, Patrícia Maria, Napoleão, Thiago Henrique, Luz Wallau, Gabriel, de -Melo-Neto, Osvaldo P., Varjal de Melo Santos, Maria Alice and Ayres, Constância Flávia Junqueira (2019) 'Polymorphisms in GSTE2 is associated with temephos resistance in Aedes aegypti'. Pesticide Biochemistry and Physiology. (In Press)

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Abstract

The glutathione S-transferases (GSTs) are enzymes involved in several distinct biological processes. In insects, the GSTs, especially delta and epsilon classes, play a key role in the metabolism of xenobiotics used to control insect populations. Here, we investigated its potential role in temephos resistance, examining the GSTE2 gene from susceptible (RecL) and resistant (RecR) strains of the mosquito Aedes aegypti, vector for several pathogenic arboviruses. Total GST enzymatic activity and the GSTE2 gene expression profile were evaluated, with the GSTE2 cDNA and genomic loci sequenced from both strains. Recombinant GSTE2 and mutants were produced in a heterologous expression system and assayed for enzyme kinetic parameters. These proteins also had their 3D structure predicted through molecular modeling. Our results showed that RecR has a profile of total GST enzymatic activity higher than RecL, with the expression of the GSTE2 gene in resistant larvae increasing six folds. Four exclusive RecR mutations were observed (L111S, I150V, E178A and A198E), which were absent in the laboratory susceptible strains. The enzymatic activity of the recombinant GSTE2 showed different kinetic parameters, with the GSTE2 RecR showing an enhanced ability to metabolize its substrate. The I150V mutation was shown to induce significant changes in catalytic parameters and a 3D modeling of GSTE2 mapped two of the RecR changes (L111S and I150V) near the enzyme's catalytic pocket, also implying an impact on its catalytic activity. Our results reinforce a potential role for GSTE2 in the metabolic resistance phenotype while contributing to the understanding of the molecular basis for the resistance mechanism.

Item Type: Article
Subjects: QU Biochemistry > Genetics > QU 450 General Works
QX Parasitology > QX 4 General works
QX Parasitology > Insects. Other Parasites > QX 510 Mosquitoes
QX Parasitology > Insects. Other Parasites > QX 525 Aedes
Faculty: Department: Biological Sciences > Vector Biology Department
Digital Object Identifer (DOI): https://doi.org/10.1016/j.pestbp.2019.10.002
Depositing User: Samantha Sheldrake
Date Deposited: 07 Nov 2019 11:50
Last Modified: 24 Jan 2020 16:46
URI: https://archive.lstmed.ac.uk/id/eprint/13030

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