UDP-glycosyltransferases (UGT), catalysing conjugation of UDP-activated sugar donors to small lipophilic chemicals, are widespread in living organisms from bacteria to fungi, plant, or animals. The progress of genome sequencing has enabled an assessment of the UGT multigene family in Haemonchus contortus (family Trichostrongylidae, Nematoda), a hematophagous gastrointestinal parasite of small ruminants. Here we report 32 putative UGT genes divided into 15 UGT families. Phylogenetic analysis in comparison with UGTs from Caenorhabditis elegans, a free-living model nematode, revealed several single member homologues, a lack of the dramatic gene expansion seen in C. elegans, but also several families (UGT365, UGT366, UGT368) expanded in H. contortus only. The assessment of constitutive UGT mRNA expression in H. contortus adults identified significant differences between females and males. In addition, we compared the expression of selected UGTs in the drug-sensitive ISE strain to two benzimidazole-resistant strains, IRE and WR, with different genetic backgrounds. Constitutive expression of UGT368B2 was significantly higher in both resistant strains than in the sensitive strain. As resistant strains were able to deactivate benzimidazole anthelmintics via glycosylation more effectively then the sensitive strain, UGT368B2 enhanced constitutive expression might contribute to drug resistance in H. contortus.

Department of Biochemical Sciences Faculty of Pharmacy Charles University Heyrovského 1203 CZ 500 05 Hradec Králové Czech Republic

Department of Pharmacology and Toxicology Faculty of Pharmacy Charles University Heyrovského 1203 CZ 500 05 Hradec Králové Czech Republic

Institute of Biodiversity Animal Health and Comparative Medicine College of Medical Veterinary and Life Sciences University of Glasgow 464 Bearsden Road Glasgow Scotland G61 1QH UK

Institute of Immunology and Microbiology 1st Faculty of Medicine Charles University Studničkova 7 CZ 128 00 Prague Czech Republic

Institute of Parasitology Slovak Academy of Sciences Hlinkova 3 SK 040 01 Košice Slovakia

Max Planck Institute for Chemical Ecology Department of Entomology Hans Knöll Straße 8 DE 07745 Jena Germany

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Ahn S.-J., Vogel H., Heckel D.G. Comparative analysis of the UDP-glycosyltransferase multigene family in insects. Insect Biochem. Mol. Biol. 2012;42:133–147. PubMed

Azzariti A., Porcelli L., Quatrale A.E., Silvestris N., Paradiso A. The coordinated role of CYP450 enzymes and P-gp in determining cancer resistance to chemotherapy. Curr. Drug Metabol. 2011;12:713–721. PubMed

Bellemare J., Rouleau M., Girard H., Harvey M., Guillemette C. Alternatively spliced products of the UGT1A gene interact with the enzymatically active proteins to inhibit glucuronosyltransferase activity in vitro. Drug Metab. Dispos. 2010;38:1785–1789. PubMed

Blackhall W.J., Prichard R.K., Beech R.N. P-glycoprotein selection in strains of Haemonchus contortus resistant to benzimidazoles. Vet. Parasitol. 2008;152:101–107. PubMed

Bock K.W. Vertebrate UDP-glucuronosyltransferases: functional and evolutionary aspects. Biochem. Pharmacol. 2003;66:691–696. PubMed

Bock K.W., Kohle C. Topological aspects of oligomeric UDP-glucuronosyltransferases in endoplasmic reticulum membranes: advances and open questions. Biochem. Pharmacol. 2009;77:1458–1465. PubMed

Bock K.W. The UDP-glycosyltransferase (UGT) superfamily expressed in humans, insects and plants: animal-plant arms-race and co-evolution. Biochem. Pharmacol. 2016;99:11–17. PubMed

Cardoso-Moreira M., Arguello J.R., Gottipati S., Harshman L.G., Grenier J.K., Clark A.G. Evidence for the fixation of gene duplications by positive selection in Drosophila. Genome Res. 2016;26:787–798. PubMed PMC

Denecke S., Fusetto R., Martelli F., Giang A., Battlay P., Fournier-Level A., O'Hair R.A., Batterham P. Multiple P450s and variation in neuronal genes underpins the response to the insecticide imidacloprid in a population of Drosophila melanogaster. Sci. Rep. 2017;7:11338. PubMed PMC

Doyle S.R., Laing R., Bartley D.J., Britton C., Chaudhry U., Gilleard J.S., Holroyd N., Mable B.K., Maitland K., Morrison A.A., Tait A., Tracey A., Berriman M., Devaney E., Cotton J.A., Sargison N.D. A genome resequencing-based genetic map reveals the recombination landscape of an outbred parasitic nematode in the presence of polyploidy and polyandry. Genome Biol Evol. 2018;10:396–409. PubMed PMC

Felsenstein J. Confidence limits on phylogenies: an approach using the bootstrap. Evolution. 1985;39:783–791. PubMed

Finel M., Kurkela M. The UDP-glucuronosyltransferases as oligomeric enzymes. Curr. Drug Metabol. 2008;9:70–76. PubMed

Fontaine P., Choe K. The transcription factor SKN-1 and detoxification gene ugt-22 alter albendazole efficacy in Caenorhabditis elegans. Int J Parasitol Drugs Drug Resist. 2018;8:312–319. PubMed PMC

Ghanizadeh H., Harrington K.C. Non-target site mechanisms of resistance to herbicides. Crit. Rev. Plant Sci. 2017;36:24–34.

Gilleard J.S., Redman E. Vol. 93. 2016. pp. 31–68. (Genetic diversity and population structure of Haemonchus contortus. Haemonchus Contortus and Haemonchosis - Past, Present and Future Trends). PubMed

Jacobs C.G., Steiger S., Heckel D.G., Wielsch N., Vilcinskas A., Vogel H. Sex, offspring and carcass determine antimicrobial peptide expression in the burying beetle. Sci. Rep. 2016;6:25409. PubMed PMC

Jones L.M., Flemming A.J., Urwin P.E. NHR-176 regulates cyp-35d1 to control hydroxylation-dependent metabolism of thiabendazole in Caenorhabditis elegans. Biochem. J. 2015;466:37–44. PubMed

Kotze A.C., Ruffell A.P., Ingham A.B. Phenobarbital induction and chemical synergism demonstrate the role of UDP-glucuronosyltransferases in detoxification of naphthalophos by Haemonchus contortus larvae. Antimicrob. Agents Chemother. 2014;58:7475–7483. PubMed PMC

Kotze A.C., Prichard R.K. Vol. 93. 2016. pp. 397–428. (Anthelmintic resistance in Haemonchus contortus: history, mechanisms and diagnosis. Haemonchus Contortus and Haemonchosis - Past, Present and Future Trends). PubMed

Kumar S., Stecher G., Li M., Knyaz C., Tamura K. MEGA X: molecular evolutionary genetics analysis across computing platforms. Mol. Biol. Evol. 2018;35:1547–1549. PubMed PMC

Kumar S., Vo A.D., Qin F.J., Li H. Comparative assessment of methods for the fusion transcripts detection from RNA-Seq data. Sci. Rep. 2016;6:21597. PubMed PMC

Kwa M.S.G., Veenstra J.G., Roos M.H. Molecular characterization of beta-tubulin genes present in benzimidazole resistant populations of Haemonchus contortus. Mol. Biochem. Parasitol. 1993;60:133–144. PubMed

Laing R., Kikuchi T., Martinelli A., Tsai I.J., Beech R.N., Redman E., Holroyd N., Bartley D.J., Beasley H., Britton C., Curran D., Devaney E., Gilabert A., Hunt M., Jackson F., Johnston S.L., Kryukov I., Li K., Morrison A.A., Reid A.J., Sargison N., Saunders G.I., Wasmuth J.D., Wolstenholme A., Berriman M., Gilleard J.S., Cotton J.A. The genome and transcriptome of Haemonchus contortus, a key model parasite for drug and vaccine discovery. Genome Biol. 2013;14:R88. PubMed PMC

Laing S.T., Ivens A., Laing R., Ravikumar S., Butler V., Woods D.J., Gilleard J.S. Characterization of the xenobiotic response of Caenorhabditis elegans to the anthelmintic drug albendazole and the identification of novel drug glucoside metabolites. Biochem. J. 2010;432:505–514. PubMed

Le S.Q., Gascuel O. An improved general amino acid replacement matrix. Mol. Biol. Evol. 2008;25:1307–1320. PubMed

Lecova L., Ruzickova M., Laing R., Vogel H., Szotakova B., Prchal L., Lamka J., Vokral I., Skalova L., Matouskova P. Reliable reference gene selection for quantitative real time PCR in Haemonchus contortus. Mol. Biochem. Parasitol. 2015;201:123–127. PubMed

Li X.X., Zhu B., Gao X.W., Liang P. Over-expression of UDP-glycosyltransferase gene UGT2B17 is involved in chlorantraniliprole resistance in Plutella xylostella (L.) Pest Manag. Sci. 2017;73:1402–1409. PubMed

Livak K.J., Schmittgen T.D. Analysis of relative gene expression data using real-time quantitative PCR and the 2(T)(-Delta Delta C) method. Methods. 2001;25:402–408. PubMed

Mackenzie P.I., Bock K.W., Burchell B., Guillemette C., Ikushiro S., Iyanagi T., Miners J.O., Owens I.S., Nebert D.W. Nomenclature update for the mammalian UDP glycosyltransferase (UGT) gene superfamily. Pharmacogenetics Genom. 2005;15:677–685. PubMed

Malik V., Black G.W. Structural, functional, and mutagenesis studies of UDP-glycosyltransferases. Structural and Mechanistic Enzymology: Bringing Together Experiments and Computing. 2012;87:87–115. PubMed

Matouskova P., Vokral I., Lamka J., Skalova L. The role of xenobiotic-metabolizing enzymes in anthelmintic deactivation and resistance in helminths. Trends Parasitol. 2016;32:481–491. PubMed

Mederos A.E., Ramos Z., Banchero G.E. First report of monepantel Haemonchus contortus resistance on sheep farms in Uruguay. Parasites Vectors. 2014;7:598. PubMed PMC

Meech R., Rogers A., Zhuang L., Lewis B.C., Miners J.O., Mackenzie P.I. Identification of residues that confer sugar selectivity to UDP-glycosyltransferase 3A (UGT3A) enzymes. J. Biol. Chem. 2012;287:24122–24130. PubMed PMC

Miley M.J., Zielinska A.K., Keenan J.E., Bratton S.M., Radominska-Pandya A., Redinbo M.R. Crystal structure of the cofactor-binding domain of the human phase II drug-metabolism enzyme UDP-glucuronosyltransferase 2B7. J. Mol. Biol. 2007;369:498–511. PubMed PMC

Petersen T.N., Brunak S., von Heijne G., Nielsen H. SignalP 4.0: discriminating signal peptides from transmembrane regions. Nat. Methods. 2011;8:785–786. PubMed

Rochat B. Role of cytochrome P450 activity in the fate of anticancer agents and in drug resistance - focus on tamoxifen, paclitaxel and imatinib metabolism. Clin. Pharmacokinet. 2005;44:349–366. PubMed

Roos M.H., Otsen M., Hoekstra R., Veenstra J.G., Lenstra J.A. Genetic analysis of inbreeding of two strains of the parasitic nematode Haemonchus contortus. Int. J. Parasitol. 2004;34:109–115. PubMed

Sievers F., Wilm A., Dineen D., Gibson T.J., Karplus K., Li W.Z., Lopez R., McWilliam H., Remmert M., Soding J., Thompson J.D., Higgins D.G. Fast, scalable generation of high-quality protein multiple sequence alignments using Clustal Omega. Mol. Syst. Biol. 2011;7:539. PubMed PMC

Stuchlíková L.R., Matoušková P., Vokřál I., Lamka J., Szotáková B., Sečkařová A., Dimunová D., Nguyen L.T., Várady M., Skálová L. Metabolism of albendazole, ricobendazole and flubendazole in Haemonchus contortus adults: sex differences, resistance-related differences and the identification of new metabolites. Int J Parasitol Drugs Drug Resist. 2018;8:50–58. PubMed PMC

Tiwari P., Sangwan R.S., Sangwan N.S. Plant secondary metabolism linked glycosyltransferases: an update on expanding knowledge and scopes. Biotechnol. Adv. 2016;34:714–739. PubMed

Untergasser A., Cutcutache I., Koressaar T., Ye J., Faircloth B.C., Remm M., Rozen S.G. Primer3-new capabilities and interfaces. Nucleic Acids Res. 2012;40:e115. PubMed PMC

Vanwyk J.A., Gerber H.M., Groeneveld H.T. A technique for the recovery of nematodes from ruminants by migration from gastrointestinal ingesta gelled in agar-large scale application Onderstepoort. J Vet Res. 1980;47:147–158. PubMed

Vogel H., Badapanda C., Knorr E., Vilcinskas A. RNA-sequencing analysis reveals abundant developmental stage-specific and immunity-related genes in the pollen beetle Meligethes aeneus. Insect Mol. Biol. 2014;23:98–112. PubMed

Vokral I., Bartikova H., Prchal L., Stuchlikova L., Skalova L., Szotakova B., Lamka J., Varady M., Kubicek V. The metabolism of flubendazole and the activities of selected biotransformation enzymes in Haemonchus contortus strains susceptible and resistant to anthelmintics. Parasitology. 2012;139:1309–1316. PubMed

Vokral I., Jirasko R., Stuchlikova L., Bartikova H., Szotakova B., Lamka J., Varady M., Skalova L. Biotransformation of albendazole and activities of selected detoxification enzymes in Haemonchus contortus strains susceptible and resistant to anthelmintics. Vet. Parasitol. 2013;196:373–381. PubMed

Williamson S.M., Storey B., Howell S., Harper K.M., Kaplan R.M., Wolstenholme A.J. Candidate anthelmintic resistance-associated gene expression and sequence polymorphisms in a triple-resistant field isolate of Haemonchus contortus. Mol. Biochem. Parasitol. 2011;180:99–105. PubMed

Yilmaz E., Ramunke S., Demeler J., Krucken J. Comparison of constitutive and thiabendazole-induced expression of five cytochrome P450 genes in fourth-stage larvae of Haemonchus contortus isolates with different drug susceptibility identifies one gene with high constitutive expression in a multi-resistant isolate. Int J Parasitol Drugs Drug Resist. 2017;7:362–369. PubMed PMC

Zuker M. Mfold web server for nucleic acid folding and hybridization prediction. Nucleic Acids Res. 2003;31:3406–3415. PubMed PMC

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