The highly polymorphic CYP6M7 cytochrome P450 gene partners with the directionally selected CYP6P9a and CYP6P9b genes to expand the pyrethroid resistance front in the malaria vector Anopheles funestus in Africa

BMC Genomics

Charles S Wondji²  Miranda Ndula²  Kayla G Barnes²  Helen Irving²  Nelson Cuamba³  Themba Mzilahowa¹  Emmanuel Chanda⁴  Sulaiman S Ibrahim²  Jacob M Riveron² 

[1] Malaria Alert Centre, College of Medicine, Blantyre, Malawi;Vector Biology Department, Liverpool School of Tropical Medicine, Pembroke place, Liverpool, L3 5QA, UK;National Institute of Health, Maputo, Mozambique;Ministry of Health, National Malaria Control Centre, Lusaka, Zambia

关键词: Directional selection;    CYP6M7;    Cytochrome P450;    Insecticide resistance;    Pyrethroids;    Malaria;    Anopheles funestus;   

Others  :  1139514
DOI  :  10.1186/1471-2164-15-817

 received in 2014-09-05, accepted in 2014-09-23,  发布年份 2014

【 摘 要 】

Background

Pyrethroid resistance in the major malaria vector Anopheles funestus is rapidly expanding across Southern Africa. It remains unknown whether this resistance has a unique origin with the same molecular basis or is multifactorial. Knowledge of the origin, mechanisms and evolution of resistance are crucial to designing successful resistance management strategies.

Results

Here, we established the resistance profile of a Zambian An. funestus population at the northern range of the resistance front. Similar to other Southern African populations, Zambian An. funestus mosquitoes are resistant to pyrethroids and carbamate, but in contrast to populations in Mozambique and Malawi, these insects are also DDT resistant. Genome-wide microarray-based transcriptional profiling and qRT-PCR revealed that the cytochrome P450 gene CYP6M7 is responsible for extending pyrethroid resistance northwards. Indeed, CYP6M7 is more over-expressed in Zambia [fold-change (FC) 37.7; 13.2 for qRT-PCR] than CYP6P9a (FC15.6; 8.9 for qRT-PCR) and CYP6P9b (FC11.9; 6.5 for qRT-PCR), whereas CYP6P9a and CYP6P9b are more highly over-expressed in Malawi and Mozambique. Transgenic expression of CYP6M7 in Drosophila melanogaster coupled with in vitro assays using recombinant enzymes and assessments of kinetic properties demonstrated that CYP6M7 is as efficient as CYP6P9a and CYP6P9b in conferring pyrethroid resistance. Polymorphism patterns demonstrate that these genes are under contrasting selection forces: the exceptionally diverse CYP6M7 likely evolves neutrally, whereas CYP6P9a and CYP6P9b are directionally selected. The higher variability of CYP6P9a and CYP6P9b observed in Zambia supports their lesser role in resistance in this country.

Conclusion

Pyrethroid resistance in Southern Africa probably has multiple origins under different evolutionary forces, which may necessitate the design of different resistance management strategies.

【 授权许可】

   
2014 Riveron et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

Figure 5.

【 参考文献 】

• [1]WHO: Malaria Report 2011. Geneva, Switzerland: World Health Organization; 2011.
• [2]WHO: Global Plan for Insecticide Resistance Management (GPIRM). Geneva, Switzerland: World Health Organization; 2012.
• [3]Okoye PN, Brooke BD, Koekemoer LL, Hunt RH, Coetzee M: Characterisation of DDT, pyrethroid and carbamate resistance in Anopheles funestus from Obuasi, Ghana. Trans R Soc Trop Med Hyg 2008, 102(6):591-598.
• [4]Riveron JM, Irving H, Ndula M, Barnes KG, Ibrahim SS, Paine MJ, Wondji CS: Directionally selected cytochrome P450 alleles are driving the spread of pyrethroid resistance in the major malaria vector Anopheles funestus. Proc Natl Acad Sci U S A 2013, 110(1):252-257.
• [5]Wondji CS, Coleman M, Kleinschmidt I, Mzilahowa T, Irving H, Ndula M, Rehman A, Morgan J, Barnes KG, Hemingway J: Impact of pyrethroid resistance on operational malaria control in Malawi. Proc Natl Acad Sci U S A 2012, 109(47):19063-19070.
• [6]Amenya DA, Naguran R, Lo TC, Ranson H, Spillings BL, Wood OR, Brooke BD, Coetzee M, Koekemoer LL: Over expression of a cytochrome P450 (CYP6P9) in a major African malaria vector, Anopheles Funestus, resistant to pyrethroids. Insect Mol Biol 2008, 17(1):19-25.
• [7]Chanda E, Hemingway J, Kleinschmidt I, Rehman AM, Ramdeen V, Phiri FN, Coetzer S, Mthembu D, Shinondo CJ, Chizema-Kawesha E, Kamuliwo M, Mukonka V, Baboo KS, Coleman M: Insecticide resistance and the future of malaria control in zambia. PLoS One 2011, 6(9):e24336.
• [8]Hunt R, Edwardes M, Coetzee M: Pyrethroid resistance in southern African Anopheles funestus extends to Likoma Island in Lake Malawi. Parasit Vectors 2010, 3:122. BioMed Central Full Text
• [9]Schlenke TA, Begun DJ: Strong selective sweep associated with a transposon insertion in Drosophila simulans. Proc Natl Acad Sci U S A 2004, 101(6):1626-1631.
• [10]Martinez-Torres D, Chandre F, Williamson MS, Darriet F, Berge JB, Devonshire AL, Guillet P, Pasteur N, Pauron D: Molecular characterization of pyrethroid knockdown resistance (kdr) in the major malaria vector Anopheles gambiae s.s. Insect Mol Biol 1998, 7(2):179-184.
• [11]Ranson H, Jensen B, Vulule JM, Wang X, Hemingway J, Collins FH: Identification of a point mutation in the voltage-gated sodium channel gene of Kenyan Anopheles gambiae associated with resistance to DDT and pyrethroids. Insect Mol Biol 2000, 9(5):491-497.
• [12]Djouaka RF, Bakare AA, Coulibaly ON, Akogbeto MC, Ranson H, Hemingway J, Strode C: 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. BMC Genomics 2008, 9:538. BioMed Central Full Text
• [13]Mitchell SN, Stevenson BJ, Muller P, Wilding CS, Egyir-Yawson A, Field SG, Hemingway J, Paine MJ, Ranson H, Donnelly MJ: Identification and validation of a gene causing cross-resistance between insecticide classes in Anopheles gambiae from Ghana. Proc Natl Acad Sci U S A 2012, 109(16):6147-6152.
• [14]Muller P, Warr E, Stevenson BJ, Pignatelli PM, Morgan JC, Steven A, Yawson AE, Mitchell SN, Ranson H, Hemingway J, Paine MJ, Donnelly MJ: Field-caught permethrin-resistant Anopheles gambiae overexpress CYP6P3, a P450 that metabolises pyrethroids. PLoS Genet 2008, 4(11):e1000286.
• [15]Gregory R, Darby AC, Irving H, Coulibaly MB, Hughes M, Koekemoer LL, Coetzee M, Ranson H, Hemingway J, Hall N, Wondji CS: A De novo expression profiling of anopheles funestus, malaria vector in Africa, using 454 pyrosequencing. PLoS One 2011, 6(2):e17418.
• [16]Crawford JE, Guelbeogo WM, Sanou A, Traore A, Vernick KD, Sagnon N, Lazzaro BP: De novo transcriptome sequencing in Anopheles funestus using Illumina RNA-seq technology. PLoS One 2010, 5(12):e14202.
• [17]Cuamba N, Morgan JC, Irving H, Steven A, Wondji CS: High level of pyrethroid resistance in an Anopheles funestus population of the Chokwe District in Mozambique. PLoS One 2010, 5(6):e11010.
• [18]Gillies MT, Coetzee M: A supplement to the Anophelinae of Africa south of the Sahara (Afrotropical region), Volume 55. South African Institute for medical research: Johannesburg; 1987.
• [19]Koekemoer LL, Kamau L, Hunt RH, Coetzee M: A cocktail polymerase chain reaction assay to identify members of the Anopheles funestus (Diptera: Culicidae) group. Am J Trop Med Hyg 2002, 66(6):804-811.
• [20]WHO: Test procedures for insecticide resistance montoring in malaria vectors, bio-efficacy and persistence of insecticides on treated surfaces. Geneva, Switzerland: World Health Organization; 1998.
• [21]Irving H, Riveron JM, Ibrahim SS, Lobo NF, Wondji CS: Positional cloning of rp2 QTL associates the P450 genes CYP6Z1, CYP6Z3 and CYP6M7 with pyrethroid resistance in the malaria vector Anopheles funestus. Heredity (Edinb) 2012, 109(6):383-392.
• [22]Wondji CS, Irving H, Morgan J, Lobo NF, Collins FH, Hunt RH, Coetzee M, Hemingway J, Ranson H: Two duplicated P450 genes are associated with pyrethroid resistance in Anopheles funestus, a major malaria vector. Genome Res 2009, 19(3):452-459.
• [23]David JP, Strode C, Vontas J, Nikou D, Vaughan A, Pignatelli PM, Louis C, Hemingway J, Ranson H: The Anopheles gambiae detoxification chip: a highly specific microarray to study metabolic-based insecticide resistance in malaria vectors. Proc Natl Acad Sci U S A 2005, 102(11):4080-4084.
• [24]Conesa A, Gotz S, Garcia-Gomez JM, Terol J, Talon M, Robles M: Blast2GO: a universal tool for annotation, visualization and analysis in functional genomics research. Bioinformatics 2005, 21(18):3674-3676.
• [25]Gotz S, Garcia-Gomez JM, Terol J, Williams TD, Nagaraj SH, Nueda MJ, Robles M, Talon M, Dopazo J, Conesa A: High-throughput functional annotation and data mining with the Blast2GO suite. Nucleic Acids Res 2008, 36(10):3420-3435.
• [26]Kwiatkowska RM, Platt N, Poupardin R, Irving H, Dabire RK, Mitchell S, Jones CM, Diabate A, Ranson H, Wondji CS: Dissecting the mechanisms responsible for the multiple insecticide resistance phenotype in Anopheles gambiae s.s., M form, from Vallee du Kou, Burkina Faso. Gene 2013, 519(1):98-106.
• [27]Schmittgen TD, Livak KJ: Analyzing real-time PCR data by the comparative C-T method. Nat Protoc 2008, 3(6):1101-1108.
• [28]McLaughlin LA, Niazi U, Bibby J, David JP, Vontas J, Hemingway J, Ranson H, Sutcliffe MJ, Paine MJ: Characterization of inhibitors and substrates of Anopheles gambiae CYP6Z2. Insect Mol Biol 2008, 17(2):125-135.
• [29]Pritchard MP, Glancey MJ, Blake JA, Gilham DE, Burchell B, Wolf CR, Friedberg T: Functional co-expression of CYP2D6 and human NADPH-cytochrome P450 reductase in Escherichia coli. Pharmacogenetics 1998, 8(1):33-42.
• [30]Stevenson BJ, Bibby J, Pignatelli P, Muangnoicharoen S, O'Neill PM, Lian LY, Muller P, Nikou D, Steven A, Hemingway J, Sutcliffe MJ, Paine MJ: Cytochrome P450 6 M2 from the malaria vector Anopheles gambiae metabolizes pyrethroids: Sequential metabolism of deltamethrin revealed. Insect Biochem Mol Biol 2011, 41(7):492-502.
• [31]Omura T, Sato R: The carbon monoxide-binding pigment of liver microsomes. I Evidence for Its Hemoprotein Nature J Biol Chem 1964, 239:2370-2378.
• [32]Strobel HW, Dignam JD: Purification and properties of NADPH-cytochrome P-450 reductase. Methods Enzymol 1978, 52:89-96.
• [33]Thompson JD, Higgins DG, Gibson TJ: CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 1994, 22(22):4673-4680.
• [34]Rozas J, Sanchez-DelBarrio JC, Messeguer X, Rozas R: DnaSP, DNA polymorphism analyses by the coalescent and other methods. Bioinformatics 2003, 19(18):2496-2497.
• [35]Tamura K, Dudley J, Nei M, Kumar S: MEGA4: molecular Evolutionary Genetics Analysis (MEGA) software version 4.0. Mol Biol Evol 2007, 24(8):1596-1599.
• [36]Clement M, Posada D, Crandall KA: TCS: a computer program to estimate gene genealogies. Mol Ecol 2000, 9(10):1657-1659.
• [37]Nei M, Gojobori T: Simple methods for estimating the numbers of synonymous and nonsynonymous nucleotide substitutions. Mol Biol Evol 1986, 3(5):418-426.
• [38]Fossog Tene B, Poupardin R, Costantini C, Awono-Ambene P, Wondji CS, Ranson H, Antonio-Nkondjio C: Resistance to DDT in an urban setting: common mechanisms implicated in both M and S forms of Anopheles gambiae in the city of Yaounde Cameroon. PLoS One 2013, 8(4):e61408.
• [39]Duangkaew P, Pethuan S, Kaewpa D, Boonsuepsakul S, Sarapusit S, Rongnoparut P: Characterization of mosquito CYP6P7 and CYP6AA3: differences in substrate preference and kinetic properties. Arch Insect Biochem Physiol 2011, 76(4):236-248.
• [40]Feyereisen R: Insect CYP Genes and P450 enzymes. In Gilbert LI. Edited by Insect Molecular Biology and Biochemistry. Oxford, UK: Elsevier; 2011:236-316.
• [41]Stevenson BJ, Pignatelli P, Nikou D, Paine MJ: Pinpointing P450s associated with pyrethroid metabolism in the dengue vector, Aedes aegypti: developing new tools to combat insecticide resistance. PLoS Negl Trop Dis 2012, 6(3):e1595.
• [42]Scollon EJ, Starr JM, Godin SJ, DeVito MJ, Hughes MF: In vitro metabolism of pyrethroid pesticides by rat and human hepatic microsomes and cytochrome p450 isoforms. Drug Metab Dispos 2009, 37(1):221-228.
• [43]Dabiré KR, Diabaté A, Namountougou M, Djogbenou L, Wondji C, Chandre F, Simard F, Ouédraogo J-B, Martin T, Weill M, Baldet T: Trends in Insecticide Resistance in Natural Populations of Malaria Vectors in Burkina Faso, West Africa: 10 Years’ Surveys. In Pest Engineering, Volume Insecticides-Pest Engineering Edited by Parveen F. 2012, 479-502. InTech
• [44]Ranson H, N'Guessan R, Lines J, Moiroux N, Nkuni Z, Corbel V: Pyrethroid resistance in African anopheline mosquitoes: what are the implications for malaria control? Trends Parasitol 2011, 27(2):91-98.
• [45]Christian RN, Strode C, Ranson H, Coetzer N, Coetzee M, Koekemoer LL: Microarray analysis of a pyrethroid resistant African malaria vector, Anopheles funestus, from southern Africa. Pestic Biochem Physiol 2011, 99(2):140-147.
• [46]Bariami V, Jones CM, Poupardin R, Vontas J, Ranson H: Gene amplification, ABC transporters and cytochrome P450s: unraveling the molecular basis of pyrethroid resistance in the dengue vector. Aedes aegypti PLoS Negl Trop Dis 2012, 6(6):e1692.
• [47]Li X, Schuler MA, Berenbaum MR: Molecular mechanisms of metabolic resistance to synthetic and natural xenobiotics. Annu Rev Entomol 2007, 52:231-253.
• [48]Zhu F, Parthasarathy R, Bai H, Woithe K, Kaussmann M, Nauen R, Harrison DA, Palli SR: A brain-specific cytochrome P450 responsible for the majority of deltamethrin resistance in the QTC279 strain of Tribolium castaneum. Proc Natl Acad Sci U S A 2010, 107(19):8557-8562.
• [49]Daborn PJ, Lumb C, Boey A, Wong W, Ffrench-Constant RH, Batterham P: Evaluating the insecticide resistance potential of eight Drosophila melanogaster cytochrome P450 genes by transgenic over-expression. Insect Biochem Mol Biol 2007, 37(5):512-519.
• [50]Ffrench-Constant RH: The molecular genetics of insecticide resistance. Genetics 2013, 194(4):807-815.
• [51]Riveron JM, Yunta C, Ibrahim SS, Djouaka R, Irving H, Menze BD, Ismail HM, Hemingway J, Ranson H, Albert A, Wondji CS: A single mutation in the GSTe2 gene allows tracking of metabolically-based insecticide resistance in a major malaria vector. Genome Biol 2014, 15(2):R27. BioMed Central Full Text
• [52]Strode C, Wondji CS, David JP, Hawkes NJ, Lumjuan N, Nelson DR, Drane DR, Karunaratne SH, Hemingway J, Black WC, Ranson H: Genomic analysis of detoxification genes in the mosquito Aedes aegypti. Insect Biochem Mol Biol 2008, 38(1):113-123.
• [53]Marcombe S, Poupardin R, Darriet F, Reynaud S, Bonnet J, Strode C, Brengues C, Yebakima A, Ranson H, Corbel V, David JP: Exploring the molecular basis of insecticide resistance in the dengue vector Aedes aegypti: a case study in Martinique Island (French West Indies). BMC Genomics 2009, 10:494. BioMed Central Full Text
• [54]Wondji CS, Hemingway J, Ranson H: Identification and analysis of Single Nucleotide Polymorphisms (SNPs) in the mosquito Anopheles funestus, malaria vector. BMC Genomics 2007, 8(1):5. BioMed Central Full Text
• [55]Wilding CS, Weetman D, Steen K, Donnelly MJ: High, clustered, nucleotide diversity in the genome of Anopheles gambiae revealed through pooled-template sequencing: implications for high-throughput genotyping protocols. BMC Genomics 2009, 10:320. BioMed Central Full Text
• [56]Rottschaefer SM, Riehle MM, Coulibaly B, Sacko M, Niare O, Morlais I, Traore SF, Vernick KD, Lazzaro BP: Exceptional diversity, maintenance of polymorphism, and recent directional selection on the APL1 malaria resistance genes of Anopheles gambiae. PLoS Biol 2011, 9(3):e1000600.
• [57]Li X, Baudry J, Berenbaum MR, Schuler MA: Structural and functional divergence of insect CYP6B proteins: from specialist to generalist cytochrome P450. Proc Natl Acad Sci U S A 2004, 101(9):2939-2944.