Expression of the cytochrome P450s, CYP6P3 and CYP6M2 are significantly elevated in multiple pyrethroid resistant populations of Anopheles gambiae s.s. from Southern Benin and Nigeria

Djouaka, Rousseau; Bakare, Adekunle A.; Coulibaly, Ousmane N; Akogbéto, Martin; Ranson, Hilary; Hemingway, Janet; Strode, Clare

doi:10.1186/1471-2164-9-538

Cited by 274 publications

(306 citation statements)

References 41 publications

Supporting

Mentioning

289

Contrasting

Order By: Relevance

“…The gene CYP6M2 was chosen for quantitative real-time PCR (qPCR) and recombinant proteinbased validation. Selection was based in part on our ability to express this class of protein in recombinant Escherichia coli systems and previous association with insecticide, although not DDT, resistance (12,16,18). The other P450 significantly overexpressed in the DDT-resistant group, CYP9L1, was not taken forward for candidate validation because only one of the four CYP9L1 gene probes reached significance and this probe could potentially crosshybridize with two other P450s, AGAP012293 and AGAP012294.…”

Section: Resultsmentioning

confidence: 99%

“…Mosquitoes were defined as resistant if they could survive a 6-h exposure to 4% DDT and a subsequent 24-h holding period. A whole-genome microarray analysis revealed that one of the most consistently overexpressed probe sets in DDT-resistant samples encoded for CYP6M2, an enzyme previously implicated in pyrethroid resistance in A. gambiae (12,16,18). Functional validation confirmed that this enzyme can metabolize both DDT and pyrethroid insecticides.…”

mentioning

confidence: 89%

“…Several studies have used microarray, quantitative trait loci, and association mapping-based approaches to identify detoxification genes, the expression of which is linked to pyrethroid resistance in the major malaria vectors (11)(12)(13)(14). Multiple candidates have been identified, with cytochrome P450s from the CYP6 class, notably CYP6P3 and CYP6M2, showing the most consistent association with pyrethroid resistance (12,15,16). Moreover, recombinant protein expression and proteomic analysis of candidates identified through these studies have confirmed an in vitro role for some of these genes in insecticide metabolism (15,17,18).…”

mentioning

confidence: 99%

See 2 more Smart Citations

Identification and validation of a gene causing cross-resistance between insecticide classes inAnopheles gambiaefrom Ghana

Mitchell

Stevenson

Müller

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

226

255

View full text Add to dashboard Cite

In the last decade there have been marked reductions in malaria incidence in sub-Saharan Africa. Sustaining these reductions will rely upon insecticides to control the mosquito malaria vectors. We report that in the primary African malaria vector, Anopheles gambiae sensu stricto, a single enzyme, CYP6M2, confers resistance to two classes of insecticide. This is unique evidence in a disease vector of cross-resistance associated with a single metabolic gene that simultaneously reduces the efficacy of two of the four classes of insecticide routinely used for malaria control. The gene-expression profile of a highly DDT-resistant population of A. gambiae s.s. from Ghana was characterized using a unique whole-genome microarray. A number of genes were significantly overexpressed compared with two susceptible West African colonies, including genes from metabolic families previously linked to insecticide resistance. One of the most significantly overexpressed probe groups (false-discovery rate-adjusted P < 0.0001) belonged to the cytochrome P450 gene CYP6M2. This gene is associated with pyrethroid resistance in wild A. gambiae s.s. populations) and can metabolize both type I and type II pyrethroids in recombinant protein assays. Using in vitro assays we show that recombinant CYP6M2 is also capable of metabolizing the organochlorine insecticide DDT in the presence of solubilizing factor sodium cholate.

show abstract

Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 89%

mentioning

confidence: 99%

See 1 more Smart Citation

Identification and validation of a gene causing cross-resistance between insecticide classes inAnopheles gambiaefrom Ghana

Mitchell

Stevenson

Müller

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

226

255

View full text Add to dashboard Cite

show abstract

“…In whole genome microarray studies of pyrethroid resistant mosquitoes, members of the cytochrome P450 class of metabolic enzymes are frequently up-regulated (Djouaka et al, 2008;Mitchell et al, 2012;Muller et al, 2008;Wondji et al, 2009) suggestive of a predominant role in the resistance phenotype. Confirmatory evidence of a direct role in resistance must come from in vitro metabolism of insecticides with heterologously expressed CYP450, and for a limited number of 4 CYP450s this evidence has been attained (Chiu et al, 2008;Mitchell et al, 2012;Muller et al, 2008;Stevenson et al, 2011).…”

Section: Introductionmentioning

confidence: 99%

A cis-regulatory sequence driving metabolic insecticide resistance in mosquitoes: Functional characterisation and signatures of selection

Wilding

Smith

Lynd

et al. 2012

Insect Biochemistry and Molecular Biology

View full text Add to dashboard Cite

Although cytochrome P450 (CYP450) enzymes are frequently up-regulated in mosquitoes resistant to insecticides, no regulatory motifs driving these expression differences with relevance to wild populations have been identified. Transposable elements (TEs) are often enriched upstream of those CYP450s involved in insecticide resistance, leading to the assumption that they contribute regulatory motifs that directly underlie the resistance phenotype. A partial CuRE1 (Culex Repetitive Element 1) transposable element is found directly upstream of CYP9M10, a cytochrome P450 implicated previously in larval resistance to permethrin in the ISOP450 strain of Cx. quinquefasciatus, but is absent from the equivalent genomic region of a susceptible strain. Via expression of CYP9M10 in E.coli we have now demonstrated time-and NADPH-dependant permethrin metabolism, prerequisites for confirmation of a role in metabolic resistance, and through qPCR shown that CYP9M10 is >20-fold over-expressed in ISOP450 compared to a susceptible strain. In a fluorescent reporter assay the region upstream of CYP9M10 from ISOP450 drove 10x expression compared to the equivalent region (lacking CuRE1) from the susceptible strain. Close correspondence with the gene expression fold-change implicates the upstream region including CuRE1 as a cis-regulatory element involved in resistance. Only a single CuRE1 bearing allele, identical to the CuRE1 bearing allele in the resistant strain, is found throughout Sub-Saharan Africa, in contrast to the diversity encountered in non-CuRE1 alleles. This suggests a single origin and subsequent spread due to selective advantage. CuRE1 is detectable using a simple diagnostic. When applied to Cx. quinquefasciatus larvae from Ghana we have demonstrated a significant association with permethrin resistance in multiple field sites (mean Odds Ratio = 3.86) suggesting this marker has relevance to natural populations of vector mosquitoes. However, when CuRE1 was excised from the allele used in the reporter assay through fusion PCR, expression was unaffected, indicating that the TE has no direct role in resistance and hence that CuRE1 is acting only as a marker of an as yet unidentified regulatory motif in the association analysis. This suggests that a re-evaluation of the assumption that TEs contribute regulatory motifs involved in gene expression may be necessary. 3 IntroductionThe control of a number of pernicious mosquito-borne diseases such as malaria, lymphatic filariasis Chanda et al., 2011;Himeidan et al., 2011; Mathias et al., 2011; Yewhalaw et al., 2011). Given that many resistance mutations are thought to be recessive in expression, development of DNA diagnostics enables disease and agricultural control programmes to identify resistance mechanisms when they are at low frequency and take ameliorating action before they cause economic or health impacts (Hemingway, 2009;Kelly-Hope et al., 2008). As shown in a number of recent studies, resistance alleles can rise towards fixation very rapidly (García et al., 2009;L...

show abstract

“…High expression of P450s has been linked to insecticide resistance in Drosophila (Sundseth et al 1990) and An. gambiae (Müller et al 2007, Djouaka et al 2008). …”

Section: Resultsmentioning

confidence: 99%

Insights into the transcriptome of oenocytes from Aedes aegypti pupae

Martins

Ramalho-Ortigão

Lobo

et al. 2011

Mem. Inst. Oswaldo Cruz

View full text Add to dashboard Cite

Expression of the cytochrome P450s, CYP6P3 and CYP6M2 are significantly elevated in multiple pyrethroid resistant populations of Anopheles gambiae s.s. from Southern Benin and Nigeria

Cited by 274 publications

References 41 publications

Identification and validation of a gene causing cross-resistance between insecticide classes inAnopheles gambiaefrom Ghana

Identification and validation of a gene causing cross-resistance between insecticide classes inAnopheles gambiaefrom Ghana

A cis-regulatory sequence driving metabolic insecticide resistance in mosquitoes: Functional characterisation and signatures of selection

Insights into the transcriptome of oenocytes from Aedes aegypti pupae

Contact Info

Product

Resources

About