title: Comparative genomics of drug resistance in Trypanosoma brucei rhodesiense.
abstract: Trypanosoma brucei rhodesiense is one of the causative agents of human sleeping sickness, a fatal disease that is transmitted by tsetse flies and restricted to Sub-Saharan Africa. Here we investigate two independent lines of T. b. rhodesiense that have been selected with the drugs melarsoprol and pentamidine over the course of 2 years, until they exhibited stable cross-resistance to an unprecedented degree. We apply comparative genomics and transcriptomics to identify the underlying mutations. Only few mutations have become fixed during selection. Three genes were affected by mutations in both lines: the aminopurine transporter AT1, the aquaporin AQP2, and the RNA-binding protein UBP1. The melarsoprol-selected line carried a large deletion including the adenosine transporter gene AT1, whereas the pentamidine-selected line carried a heterozygous point mutation in AT1, G430R, which rendered the transporter non-functional. Both resistant lines had lost AQP2, and both lines carried the same point mutation, R131L, in the RNA-binding motif of UBP1. The finding that concomitant deletion of the known resistance genes AT1 and AQP2 in T. b. brucei failed to phenocopy the high levels of resistance of the T. b. rhodesiense mutants indicated a possible role of UBP1 in melarsoprol-pentamidine cross-resistance. However, homozygous in situ expression of UBP1-Leu(131) in T. b. brucei did not affect the sensitivity to melarsoprol or pentamidine.
authors.affiliation: Swiss Tropical and Public Health Institute, Socinstrasse 57, 4051, Basel, Switzerland.|University of Basel, 4000, Basel, Switzerland.
authors.forename: Fabrice E
authors.initials: FE
authors.lastname: Graf
authors.affiliation: Swiss Tropical and Public Health Institute, Socinstrasse 57, 4051, Basel, Switzerland.|University of Basel, 4000, Basel, Switzerland.
authors.forename: Philipp
authors.initials: P
authors.lastname: Ludin
authors.affiliation: Swiss Tropical and Public Health Institute, Socinstrasse 57, 4051, Basel, Switzerland.|University of Basel, 4000, Basel, Switzerland.
authors.forename: Christian
authors.initials: C
authors.lastname: Arquint
authors.affiliation: Swiss Tropical and Public Health Institute, Socinstrasse 57, 4051, Basel, Switzerland.|University of Basel, 4000, Basel, Switzerland.
authors.forename: Remo S
authors.initials: RS
authors.lastname: Schmidt
authors.affiliation: Swiss Tropical and Public Health Institute, Socinstrasse 57, 4051, Basel, Switzerland.|University of Basel, 4000, Basel, Switzerland.
authors.forename: Nadia
authors.initials: N
authors.lastname: Schaub
authors.affiliation: Swiss Tropical and Public Health Institute, Socinstrasse 57, 4051, Basel, Switzerland.|University of Basel, 4000, Basel, Switzerland.
authors.forename: Christina
authors.initials: C
authors.lastname: Kunz Renggli
authors.affiliation: Institute of Infection, Immunity and Inflammation, University of Glasgow, Glasgow, G12 8TA, UK.
authors.forename: Jane C
authors.initials: JC
authors.lastname: Munday
authors.affiliation: Institute of Infection, Immunity and Inflammation, University of Glasgow, Glasgow, G12 8TA, UK.
authors.forename: Jessica
authors.initials: J
authors.lastname: Krezdorn
authors.affiliation: Biological Chemistry and Drug Discovery, School of Life Sciences, University of Dundee, Dow Street, Dundee, DD1 5EH, UK.|The University of Kent, Canterbury, Kent, CT2 7NZ, UK.
authors.forename: Nicola
authors.initials: N
authors.lastname: Baker
authors.affiliation: Biological Chemistry and Drug Discovery, School of Life Sciences, University of Dundee, Dow Street, Dundee, DD1 5EH, UK.
authors.forename: David
authors.initials: D
authors.lastname: Horn
authors.affiliation: Swiss Tropical and Public Health Institute, Socinstrasse 57, 4051, Basel, Switzerland.|University of Basel, 4000, Basel, Switzerland.
authors.forename: Oliver
authors.initials: O
authors.lastname: Balmer
authors.affiliation: Department of Ecology and Evolutionary Biology, Yale University, New Haven, CT, USA.
authors.forename: Adalgisa
authors.initials: A
authors.lastname: Caccone
authors.affiliation: Institute of Infection, Immunity and Inflammation, University of Glasgow, Glasgow, G12 8TA, UK.
authors.forename: Harry P
authors.initials: HP
authors.lastname: de Koning
authors.affiliation: Swiss Tropical and Public Health Institute, Socinstrasse 57, 4051, Basel, Switzerland. pascal.maeser@unibas.ch.|University of Basel, 4000, Basel, Switzerland. pascal.maeser@unibas.ch.
authors.forename: Pascal
authors.identifier: 0000-0003-3122-1941
authors.initials: P
authors.lastname: Mäser
mesh_terms: D000595:Amino Acid Sequence; D020346:Aquaporins / Q000235:genetics / Q000378:metabolism; D055028:Comparative Genomic Hybridization; D016054:DNA, Protozoan / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D004351:Drug Resistance / Q000235:genetics*; D018503:Genome, Protozoan; D006579:Heterozygote; D006801:Humans; D008297:Male; D008549:Melarsoprol / Q000494:pharmacology; D033703:Nucleoside Transport Proteins / Q000235:genetics / Q000378:metabolism; D021261:Parasitic Sensitivity Tests; D010419:Pentamidine / Q000494:pharmacology; D010641:Phenotype; D020641:Polymorphism, Single Nucleotide; D015800:Protozoan Proteins / Q000235:genetics / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D014344:Trypanocidal Agents / Q000494:pharmacology; D016833:Trypanosoma brucei rhodesiense / Q000187:drug effects / Q000235:genetics* / Q000302:isolation & purification; D014353:Trypanosomiasis, African / Q000175:diagnosis / Q000469:parasitology
chemical_list: D020346:Aquaporins; D016054:DNA, Protozoan; D033703:Nucleoside Transport Proteins; D015800:Protozoan Proteins; D016601:RNA-Binding Proteins; D014344:Trypanocidal Agents; D010419:Pentamidine; D008549:Melarsoprol
keywords: African trypanosomes; Aquaporin; Melarsoprol; Pentamidine; Purine permease; RNA-binding protein