title: Artemisinin resistance in rodent malaria--mutation in the AP2 adaptor μ-chain suggests involvement of endocytosis and membrane protein trafficking.
abstract: BACKGROUND
  The control of malaria, caused by Plasmodium falciparum, is hampered by the relentless evolution of drug resistance. Because artemisinin derivatives are now used in the most effective anti-malarial therapy, resistance to artemisinin would be catastrophic. Indeed, studies suggest that artemisinin resistance has already appeared in natural infections. Understanding the mechanisms of resistance would help to prolong the effective lifetime of these drugs. Genetic markers of resistance are therefore required urgently. Previously, a mutation in a de-ubiquitinating enzyme was shown to confer artemisinin resistance in the rodent malaria parasite Plasmodium chabaudi.
  
  
  METHODS
  Here, for a mutant P. chabaudi malaria parasite and its immediate progenitor, the in vivo artemisinin resistance phenotypes and the mutations arising using Illumina whole-genome re-sequencing were compared.
  
  
  RESULTS
  An increased artemisinin resistance phenotype is accompanied by one non-synonymous substitution. The mutated gene encodes the μ-chain of the AP2 adaptor complex, a component of the endocytic machinery. Homology models indicate that the mutated residue interacts with a cargo recognition sequence. In natural infections of the human malaria parasite P. falciparum, 12 polymorphisms (nine SNPs and three indels) were identified in the orthologous gene.
  
  
  CONCLUSIONS
  An increased artemisinin-resistant phenotype occurs along with a mutation in a functional element of the AP2 adaptor protein complex. This suggests that endocytosis and trafficking of membrane proteins may be involved, generating new insights into possible mechanisms of resistance. The genotypes of this adaptor protein can be evaluated for its role in artemisinin responses in human infections of P. falciparum.
authors.affiliation: Centro de Malaria & Doenças Tropicais,LA/IHMT/Universidade Nova de Lisboa, Lisbon, Portugal. gisela.henriques@lshtm.ac.uk.
authors.forename: Gisela
authors.initials: G
authors.lastname: Henriques
authors.forename: Axel
authors.initials: A
authors.lastname: Martinelli
authors.forename: Louise
authors.initials: L
authors.lastname: Rodrigues
authors.forename: Katarzyna
authors.initials: K
authors.lastname: Modrzynska
authors.forename: Richard
authors.initials: R
authors.lastname: Fawcett
authors.forename: Douglas R
authors.initials: DR
authors.lastname: Houston
authors.forename: Sofia T
authors.initials: ST
authors.lastname: Borges
authors.forename: Umberto
authors.initials: U
authors.lastname: d'Alessandro
authors.forename: Halidou
authors.initials: H
authors.lastname: Tinto
authors.forename: Corine
authors.initials: C
authors.lastname: Karema
authors.forename: Paul
authors.initials: P
authors.lastname: Hunt
authors.forename: Pedro
authors.initials: P
authors.lastname: Cravo
mesh_terms: D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000818:Animals; D000962:Antimalarials / Q000494:pharmacology*; D037621:Artemisinins / Q000494:pharmacology*; D004252:DNA Mutational Analysis; D004351:Drug Resistance; D005819:Genetic Markers; D006801:Humans; D051379:Mice; D008808:Mice, Inbred CBA; D008969:Molecular Sequence Data; D020125:Mutation, Missense; D016788:Plasmodium chabaudi / Q000187:drug effects* / Q000235:genetics* / Q000302:isolation & purification; D010963:Plasmodium falciparum / Q000187:drug effects / Q000235:genetics / Q000302:isolation & purification; D011487:Protein Conformation; D015800:Protozoan Proteins / Q000737:chemistry / Q000235:genetics*
chemical_list: D000962:Antimalarials; D037621:Artemisinins; D005819:Genetic Markers; D015800:Protozoan Proteins; C031327:artemisinin
references.citation: Science. 2012 Apr 6;336(6077):79-82
references.pmid: 22491853
references.citation: Antimicrob Agents Chemother. 2006 Feb;50(2):480-9
references.pmid: 16436700
references.citation: Trends Parasitol. 2001 May;17(5):236-42
references.pmid: 11323308
references.citation: Mol Biochem Parasitol. 2002 Aug 28;123(2):95-104
references.pmid: 12270625
references.citation: N Engl J Med. 2009 Jul 30;361(5):455-67
references.pmid: 19641202
references.citation: Genome Res. 2008 Nov;18(11):1851-8
references.pmid: 18714091
references.citation: Genetics. 2007 Jul;176(3):1691-701
references.pmid: 17507681
references.citation: Malar J. 2009 Jan 12;8:10
references.pmid: 19138388
references.citation: Genome Res. 2001 Oct;11(10):1725-9
references.pmid: 11591649
references.citation: Antimicrob Agents Chemother. 2008 May;52(5):1840-2
references.pmid: 18316523
references.citation: Mol Microbiol. 2007 Jul;65(1):27-40
references.pmid: 17581118
references.citation: N Engl J Med. 2008 Dec 11;359(24):2619-20
references.pmid: 19064625
references.citation: Nature. 1976 Jun 17;261(5561):585-6
references.pmid: 934297
references.citation: Antimicrob Agents Chemother. 2010 May;54(5):1872-7
references.pmid: 20160056
references.citation: Traffic. 2010 Feb;11(2):236-49
references.pmid: 20015114
references.citation: Nature. 2010 Oct 28;467(7319):1061-73
references.pmid: 20981092
references.citation: Lancet. 2012 May 26;379(9830):1960-6
references.pmid: 22484134
references.citation: Exp Parasitol. 1981 Dec;52(3):419-26
references.pmid: 6947894
references.citation: Antimicrob Agents Chemother. 2008 Jan;52(1):98-109
references.pmid: 17938190
references.citation: Trends Parasitol. 2011 Feb;27(2):73-81
references.pmid: 21169061
references.citation: Nat Protoc. 2010 Apr;5(4):725-38
references.pmid: 20360767
references.citation: N Engl J Med. 2009 Jul 30;361(5):540-1
references.pmid: 19641219
references.citation: J Cell Sci. 2010 Feb 1;123(Pt 3):441-50
references.pmid: 20067995
references.citation: Lancet. 2005 Dec 3;366(9501):1960-3
references.pmid: 16325698
references.citation: Drug Dev Res. 2010 Feb;71(1):12-19
references.pmid: 21399699
references.citation: J Infect Dis. 2010 Nov 1;202(9):1362-8
references.pmid: 20863228
references.citation: Genome Biol. 2007;8(9):R180
references.pmid: 17784950
references.citation: Cell Microbiol. 2008 Feb;10(2):452-64
references.pmid: 17944961
references.citation: Antimicrob Agents Chemother. 2003 Feb;47(2):709-18
references.pmid: 12543682
references.citation: Blood. 2010 Mar 25;115(12):2500-7
references.pmid: 20093402
references.citation: Acta Trop. 1993 Sep;54(3-4):237-50
references.pmid: 7902661
references.citation: Curr Biol. 2001 Jun 5;11(11):896-900
references.pmid: 11516654
references.citation: Molecules. 2010 Mar 12;15(3):1705-21
references.pmid: 20336009
references.citation: Trop Med Int Health. 2008 Feb;13(2):199-207
references.pmid: 18304266
references.citation: Biochem Pharmacol. 2009 Feb 1;77(3):322-36
references.pmid: 19022224
references.citation: Lancet. 2004 Jul 31-Aug 6;364(9432):438-447
references.pmid: 15288742
references.citation: Nat Struct Mol Biol. 2005 Jul;12(7):628-9
references.pmid: 15937493
references.citation: Antimicrob Agents Chemother. 2004 Jul;48(7):2370-8
references.pmid: 15215083
references.citation: Mol Biol Cell. 1999 Nov;10(11):3643-59
references.pmid: 10564262
references.citation: Proc Natl Acad Sci U S A. 2011 Jul 12;108(28):11405-10
references.pmid: 21709259
references.citation: Am J Trop Med Hyg. 1995 Jun;52(6):565-8
references.pmid: 7611566
references.citation: Nature. 2003 Aug 21;424(6951):957-61
references.pmid: 12931192
references.citation: Nature. 2000 Feb 24;403(6772):906-9
references.pmid: 10706290
references.citation: Antimicrob Agents Chemother. 2010 Jul;54(7):2886-92
references.pmid: 20421395
references.citation: PLoS Negl Trop Dis. 2012;6(2):e1512
references.pmid: 22348164
references.citation: Proc Natl Acad Sci U S A. 2013 Jan 2;110(1):240-5
references.pmid: 23248304
references.citation: Mol Biochem Parasitol. 2002 Jul;122(2):217-21
references.pmid: 12106876
references.citation: Int J Parasitol. 2011 Feb;41(2):165-71
references.pmid: 20858498
references.citation: Annu Rev Biochem. 2003;72:395-447
references.pmid: 12651740
references.citation: Parasitology. 1975 Feb;70(1):19-24
references.pmid: 1118185
references.citation: Annu Rev Cell Dev Biol. 2004;20:153-91
references.pmid: 15473838
references.citation: BMC Genomics. 2012 Mar 21;13:106
references.pmid: 22435897
references.citation: BMC Genomics. 2010 Sep 16;11:499
references.pmid: 20846421
references.citation: Antimicrob Agents Chemother. 2011 Jan;55(1):50-5
references.pmid: 21041511
references.citation: Trop Med Int Health. 2007 Mar;12(3):353-62
references.pmid: 17313506
references.citation: Antimicrob Agents Chemother. 2010 Jun;54(6):2455-64
references.pmid: 20350946
references.citation: FEBS J. 2007 Sep;274(18):4688-98
references.pmid: 17824955
references.citation: Science. 2007 Jan 12;315(5809):201-5
references.pmid: 17218518
references.citation: Nat Rev Mol Cell Biol. 2009 Sep;10(9):583-96
references.pmid: 19696796
references.citation: Bioinformatics. 2009 Aug 15;25(16):2078-9
references.pmid: 19505943
references.citation: Trans R Soc Trop Med Hyg. 2006 Jun;100(6):509-14
references.pmid: 16337665
references.citation: Nature. 2012 Jan 25;482(7384):232-6
references.pmid: 22278056
references.citation: EMBO J. 1999 Jul 15;18(14):3897-908
references.pmid: 10406795
references.citation: Biochem Pharmacol. 2011 Aug 1;82(3):236-47
references.pmid: 21596024
references.citation: Cell. 2010 Jun 25;141(7):1220-9
references.pmid: 20603002
references.citation: Cell. 2002 May 17;109(4):523-35
references.pmid: 12086608
references.citation: J Infect Dis. 2006 Aug 15;194(4):528-35
references.pmid: 16845638
references.citation: Malar J. 2009 Jan 12;8:11
references.pmid: 19138391
references.citation: Antimicrob Agents Chemother. 2011 Oct;55(10):4858-65
references.pmid: 21709099