title: UDP-galactose and acetyl-CoA transporters as Plasmodium multidrug resistance genes.
abstract: A molecular understanding of drug resistance mechanisms enables surveillance of the effectiveness of new antimicrobial therapies during development and deployment in the field. We used conventional drug resistance selection as well as a regime of limiting dilution at early stages of drug treatment to probe two antimalarial imidazolopiperazines, KAF156 and GNF179. The latter approach permits the isolation of low-fitness mutants that might otherwise be out-competed during selection. Whole-genome sequencing of 24 independently derived resistant Plasmodium falciparum clones revealed four parasites with mutations in the known cyclic amine resistance locus (pfcarl) and a further 20 with mutations in two previously unreported P. falciparum drug resistance genes, an acetyl-CoA transporter (pfact) and a UDP-galactose transporter (pfugt). Mutations were validated both in vitro by CRISPR editing in P. falciparum and in vivo by evolution of resistant Plasmodium berghei mutants. Both PfACT and PfUGT were localized to the endoplasmic reticulum by fluorescence microscopy. As mutations in pfact and pfugt conveyed resistance against additional unrelated chemical scaffolds, these genes are probably involved in broad mechanisms of antimalarial drug resistance.
authors.affiliation: Novartis Institute for Tropical Diseases, 138670 Singapore.|Department of Pharmacy, Faculty of Science, National University of Singapore, 119077 Singapore.
authors.forename: Michelle Yi-Xiu
authors.initials: MY
authors.lastname: Lim
authors.affiliation: Department of Pediatrics, School of Medicine, University of California, San Diego, La Jolla, California 92093, USA.
authors.forename: Gregory
authors.initials: G
authors.lastname: LaMonte
authors.affiliation: Department of Microbiology & Immunology, Columbia University Medical Center, New York, New York 10032, USA.|Malaria Programme, Wellcome Trust Sanger Institute, Wellcome Genome Campus, Cambridge CB10 1SA, UK.
authors.forename: Marcus C S
authors.initials: MCS
authors.lastname: Lee
authors.affiliation: Department of Pediatrics, School of Medicine, University of California, San Diego, La Jolla, California 92093, USA.
authors.forename: Christin
authors.initials: C
authors.lastname: Reimer
authors.affiliation: Novartis Institute for Tropical Diseases, 138670 Singapore.
authors.forename: Bee Huat
authors.initials: BH
authors.lastname: Tan
authors.affiliation: Department of Pediatrics, School of Medicine, University of California, San Diego, La Jolla, California 92093, USA.
authors.forename: Victoria
authors.initials: V
authors.lastname: Corey
authors.affiliation: Novartis Institute for Tropical Diseases, 138670 Singapore.|Department of Microbiology & Immunology, Yong Loo Lin School of Medicine, National University of Singapore, National University Health System, 117545 Singapore.
authors.forename: Bianca F
authors.initials: BF
authors.lastname: Tjahjadi
authors.affiliation: Novartis Institute for Tropical Diseases, 138670 Singapore.
authors.forename: Adeline
authors.initials: A
authors.lastname: Chua
authors.affiliation: Department of Pediatrics, School of Medicine, University of California, San Diego, La Jolla, California 92093, USA.
authors.forename: Marie
authors.initials: M
authors.lastname: Nachon
authors.affiliation: Laboratory of Biopolymers and Supramolecular Nanomaterials, Faculty of Pharmacy, Université Libre de Bruxelles, Brussels, Belgium.
authors.forename: René
authors.initials: R
authors.lastname: Wintjens
authors.affiliation: Novartis Institute for Tropical Diseases, 138670 Singapore.
authors.forename: Peter
authors.initials: P
authors.lastname: Gedeck
authors.affiliation: Department of Microbiology & Immunology, Yong Loo Lin School of Medicine, National University of Singapore, National University Health System, 117545 Singapore.|Singapore Immunology Network (SIgN), A*Star, 138648 Singapore.
authors.forename: Benoit
authors.initials: B
authors.lastname: Malleret
authors.affiliation: Singapore Immunology Network (SIgN), A*Star, 138648 Singapore.
authors.forename: Laurent
authors.identifier: http://orcid.org/0000-0003-0349-1557
authors.initials: L
authors.lastname: Renia
authors.affiliation: Novartis Institute for Tropical Diseases, 138670 Singapore.
authors.forename: Ghislain M C
authors.initials: GMC
authors.lastname: Bonamy
authors.affiliation: Department of Pharmacy, Faculty of Science, National University of Singapore, 119077 Singapore.
authors.forename: Paul Chi-Lui
authors.initials: PC
authors.lastname: Ho
authors.affiliation: Novartis Institute for Tropical Diseases, 138670 Singapore.
authors.forename: Bryan K S
authors.initials: BKS
authors.lastname: Yeung
authors.affiliation: Center for Advanced Technology, Department of Biochemistry and Biophysics, University of California, San Francisco, California 94143, USA.
authors.forename: Eric D
authors.identifier: http://orcid.org/0000-0001-8079-918X
authors.initials: ED
authors.lastname: Chow
authors.affiliation: Novartis Institute for Tropical Diseases, 138670 Singapore.
authors.forename: Liting
authors.initials: L
authors.lastname: Lim
authors.affiliation: Department of Microbiology & Immunology, Columbia University Medical Center, New York, New York 10032, USA.|Division of Infectious Diseases, Department of Medicine, Columbia University Medical Center, New York, New York 10032, USA.
authors.forename: David A
authors.initials: DA
authors.lastname: Fidock
authors.affiliation: Novartis Institute for Tropical Diseases, 138670 Singapore.|Department of Microbiology & Immunology, Yong Loo Lin School of Medicine, National University of Singapore, National University Health System, 117545 Singapore.
authors.forename: Thierry T
authors.initials: TT
authors.lastname: Diagana
authors.affiliation: Department of Pediatrics, School of Medicine, University of California, San Diego, La Jolla, California 92093, USA.
authors.forename: Elizabeth A
authors.initials: EA
authors.lastname: Winzeler
authors.affiliation: Novartis Institute for Tropical Diseases, 138670 Singapore.|Department of Microbiology & Immunology, Yong Loo Lin School of Medicine, National University of Singapore, National University Health System, 117545 Singapore.
authors.forename: Pablo
authors.initials: P
authors.lastname: Bifani