Profiling of abundant proteins associated with dichlorodiphenyltrichloroethane resistance in Drosophila melanogaster

Abstract

Dichlorodiphenyltrichloroethane (DDT) metabolism-based resistance in Drosophila melanogaster is a complex metabolic system associated with the transcription of detoxification related genes, ion transport, lipid and sugar metabolism pathways. However, little is known about the differences regarding the proteome of field- and laboratory-selected resistant Drosophila genotypes. We investigated the impact of DDT resistance in the abundant proteome of field- and laboratory- selected resistant Drosophila using a two-dimensional gel electrophoresis DDT reference map. Proteomic profiling was performed in two DDT susceptible genotypes (Canton-S and 91-C) and three DDT resistant lines (Rst(2)DDT(91-R), Rst(2)DDT(Wisconsin) and Rst(2)DDT(Hikone-R)). Protein spots were stained with Coomassie blue and compared using PDQuest software. Selected protein spots were cut out and analyzed using matrix assisted laser desorption-time of flight mass spectrometry. Querying the NCBInr. 10.21.2003 database with mass spectrometric data yielded the identity of 21 differentially translated proteins in Rst(2)DDT(91-R), Rst(2)DDT(Wisconsin) and Canton-S representing proteins putatively involved in biochemical pathways such as glycolysis and gluconeogenesis, the pentose phosphate pathway, the Krebs cycle and fatty acid oxidation. We hypothesize that both strategies are aimed to use of the pentose phosphate pathway to increase glucose utilization while Rst(2)DDT(91-R) relies primarily on glycolysis to produce reduced NADP and increase DDT detoxification. DDT exposure in Canton-S induced six proteins, while four proteins were repressed in Rst(2)DDT(Hikone-R). Our data suggest that insecticide resistance appears to impact different metabolic pathways in Drosophila genotypes selected with the same pesticide (DDT).

Genome-Wide Sequencing and an Open Reading Frame Analysis of Dichlorodiphenyltrichloroethane (DDT) Susceptible (91-C) and Resistant (91-R) Drosophila melanogaster Laboratory Populations

“…In addition to CYP6G1 , Festucci-Buselli et al [32] identified CYP12D1 as conferring resistance to DDT. The only gene identified in the literature search that overlapped with this study was Nina C , a gene with kinase activity and related to sensory transduction/vision, which was shown by Pedra et al [31] to be overexpressed in DDT resistant strains and shown in this study to contain structural changes. Literature examining structural changes revealed two genes, CYP6A2 and para , to contain mutations within their coding sequence (Table 5).…”

“…The current work adds one more critical aspect to our understanding of the molecular changes that have occurred under DDT selection as previous work respectively defined genome-wide and partially proteome-wide differential expression of transcripts and proteins [30], [31]. With a total number of resistant SNPs/DIPs of 710, and 565 of these being novel mutations, spanning across all four chromosomes, it highlights that selection with DDT may result, on average, with the selection for novel mutations, potentially with some or many of these being associated with or directly involved in DDT resistance (Figure 2).…”

Genome‐wide analysis of phenobarbital‐inducible genes in Drosophila melanogaster

“…1b). Additionally, two‐dimensional gel comparisons between DDT resistant and susceptible Drosophila protein patterns revealed differential expression of proteins associated with carbohydrate and lipid metabolism (Pedra et al ., 2005). In insects DDT metabolism has been shown to be associated with glucose utilization (Plapp, 1970; Maltseva & Golovleva, 1982) and in mammals DDT has also been shown to affect β‐oxidation of fatty acids (Hinton et al ., 1978; Oda et al ., 1994).…”

Quantitative Trait Loci Mapping of Genome Regions Controlling Permethrin Resistance in the Mosquito Aedes aegypti

“…Limitations associated with the use of targeted microarray chips are that only those genes printed on the chip can be examined and only those genes whose function is related to changes in transcriptional activity can be identified. While proteomic approaches toward identifying resistant genes in Drosophila melanogaster have been developed (Pedra et al 2005), similar tools have not been applied in mosquitoes.…”