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EMBO J. Aug 1992; 11(8): 3067–3075.
PMCID: PMC556790

Selection for high-level chloroquine resistance results in deamplification of the pfmdr1 gene and increased sensitivity to mefloquine in Plasmodium falciparum.

Abstract

A chloroquine resistant cloned isolate of Plasmodium falciparum, FAC8, which carries an amplification in the pfmdr1 gene was selected for high-level chloroquine resistance, resulting in a cell line resistant to a 10-fold higher concentration of chloroquine. These cells were found to have lost the amplification in pfmdr1 and to no longer over-produce the protein product termed P-glycoprotein homologue 1 (Pgh1). The pfmdr1 gene from this highly resistant cell line was not found to encode any amino acid changes that would account for increased resistance. Verapamil, which reverses chloroquine resistance in FAC8, also reversed high-level chloroquine resistance. Furthermore, verapamil caused a biphasic reversal of chloroquine resistance as the high-level resistance was very sensitive to low amounts of verapamil. These data suggest that over-expression of the P-glycoprotein homologue is incompatible with high levels of chloroquine resistance. In order to show that these results were applicable to other chloroquine selected lines, two additional mutants were selected for resistance to high levels of chloroquine. In both cases they were found to deamplify pfmdr1. Interestingly, while the level of chloroquine resistance of these mutants increased, they became more sensitive to mefloquine. This suggests a linkage between the copy number of the pfmdr1 gene and the level of chloroquine and mefloquine resistance.

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  • Peterson DS, Walliker D, Wellems TE. Evidence that a point mutation in dihydrofolate reductase-thymidylate synthase confers resistance to pyrimethamine in falciparum malaria. Proc Natl Acad Sci U S A. 1988 Dec;85(23):9114–9118. [PMC free article] [PubMed]
  • Saiki RK, Gelfand DH, Stoffel S, Scharf SJ, Higuchi R, Horn GT, Mullis KB, Erlich HA. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science. 1988 Jan 29;239(4839):487–491. [PubMed]
  • Trager W, Jenson JB. Cultivation of malarial parasites. Nature. 1978 Jun 22;273(5664):621–622. [PubMed]
  • Triglia T, Foote SJ, Kemp DJ, Cowman AF. Amplification of the multidrug resistance gene pfmdr1 in Plasmodium falciparum has arisen as multiple independent events. Mol Cell Biol. 1991 Oct;11(10):5244–5250. [PMC free article] [PubMed]
  • Warhurst DC. Antimalarial schizontocides: why a permease is necessary. Parasitol Today. 1986 Dec;2(12):331–334. [PubMed]
  • Wellems TE, Panton LJ, Gluzman IY, do Rosario VE, Gwadz RW, Walker-Jonah A, Krogstad DJ. Chloroquine resistance not linked to mdr-like genes in a Plasmodium falciparum cross. Nature. 1990 May 17;345(6272):253–255. [PubMed]
  • Wilson CM, Serrano AE, Wasley A, Bogenschutz MP, Shankar AH, Wirth DF. Amplification of a gene related to mammalian mdr genes in drug-resistant Plasmodium falciparum. Science. 1989 Jun 9;244(4909):1184–1186. [PubMed]
  • Bianco AE, Battye FL, Brown GV. Plasmodium falciparum: rapid quantification of parasitemia in fixed malaria cultures by flow cytometry. Exp Parasitol. 1986 Oct;62(2):275–282. [PubMed]
  • Bianco AE, Favaloro JM, Burkot TR, Culvenor JG, Crewther PE, Brown GV, Anders RF, Coppel RL, Kemp DJ. A repetitive antigen of Plasmodium falciparum that is homologous to heat shock protein 70 of Drosophila melanogaster. Proc Natl Acad Sci U S A. 1986 Nov;83(22):8713–8717. [PMC free article] [PubMed]
  • Biggs BA, Kemp DJ, Brown GV. Subtelomeric chromosome deletions in field isolates of Plasmodium falciparum and their relationship to loss of cytoadherence in vitro. Proc Natl Acad Sci U S A. 1989 Apr;86(7):2428–2432. [PMC free article] [PubMed]
  • Coppel RL, Culvenor JG, Bianco AE, Crewther PE, Stahl HD, Brown GV, Anders RF, Kemp DJ. Variable antigen associated with the surface of erythrocytes infected with mature stages of Plasmodium falciparum. Mol Biochem Parasitol. 1986 Sep;20(3):265–277. [PubMed]
  • Corcoran LM, Forsyth KP, Bianco AE, Brown GV, Kemp DJ. Chromosome size polymorphisms in Plasmodium falciparum can involve deletions and are frequent in natural parasite populations. Cell. 1986 Jan 17;44(1):87–95. [PubMed]
  • Corcoran LM, Thompson JK, Walliker D, Kemp DJ. Homologous recombination within subtelomeric repeat sequences generates chromosome size polymorphisms in P. falciparum. Cell. 1988 Jun 3;53(5):807–813. [PubMed]
  • Cowman AF, Morry MJ, Biggs BA, Cross GA, Foote SJ. Amino acid changes linked to pyrimethamine resistance in the dihydrofolate reductase-thymidylate synthase gene of Plasmodium falciparum. Proc Natl Acad Sci U S A. 1988 Dec;85(23):9109–9113. [PMC free article] [PubMed]
  • Cowman AF, Karcz S, Galatis D, Culvenor JG. A P-glycoprotein homologue of Plasmodium falciparum is localized on the digestive vacuole. J Cell Biol. 1991 Jun;113(5):1033–1042. [PMC free article] [PubMed]
  • Draper CC, Hills M, Kilimali VA, Brubaker G. Serial studies on the evolution of drug resistance in malaria in an area of east Africa: findings from 1979 up to 1986. J Trop Med Hyg. 1988 Oct;91(5):265–273. [PubMed]
  • Endicott JA, Ling V. The biochemistry of P-glycoprotein-mediated multidrug resistance. Annu Rev Biochem. 1989;58:137–171. [PubMed]
  • Feinberg AP, Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. [PubMed]
  • Foote SJ, Thompson JK, Cowman AF, Kemp DJ. Amplification of the multidrug resistance gene in some chloroquine-resistant isolates of P. falciparum. Cell. 1989 Jun 16;57(6):921–930. [PubMed]
  • Foote SJ, Kyle DE, Martin RK, Oduola AM, Forsyth K, Kemp DJ, Cowman AF. Several alleles of the multidrug-resistance gene are closely linked to chloroquine resistance in Plasmodium falciparum. Nature. 1990 May 17;345(6272):255–258. [PubMed]
  • Geary TG, Jensen JB, Ginsburg H. Uptake of [3H]chloroquine by drug-sensitive and -resistant strains of the human malaria parasite Plasmodium falciparum. Biochem Pharmacol. 1986 Nov 1;35(21):3805–3812. [PubMed]
  • Ginsburg H, Stein WD. Kinetic modelling of chloroquine uptake by malaria-infected erythrocytes. Assessment of the factors that may determine drug resistance. Biochem Pharmacol. 1991 May 15;41(10):1463–1470. [PubMed]
  • Krogstad DJ, Schlesinger PH. The basis of antimalarial action: non-weak base effects of chloroquine on acid vesicle pH. Am J Trop Med Hyg. 1987 Mar;36(2):213–220. [PubMed]
  • Krogstad DJ, Gluzman IY, Kyle DE, Oduola AM, Martin SK, Milhous WK, Schlesinger PH. Efflux of chloroquine from Plasmodium falciparum: mechanism of chloroquine resistance. Science. 1987 Nov 27;238(4831):1283–1285. [PubMed]
  • Limpaiboon T, Shirley MW, Kemp DJ, Saul A. 7H8/6, a multicopy DNA probe for distinguishing isolates of Plasmodium falciparum. Mol Biochem Parasitol. 1991 Aug;47(2):197–206. [PubMed]
  • MABERTI S. [The development of resistance to pyrimethamine. Presentation of 15 cases studied in Trujillo, Venezuela]. Arch Venez Med Trop Parasitol Med. 1960 Dec;3:239–259. [PubMed]
  • Makler MT, Lee LG, Recktenwald D. Thiazole orange: a new dye for Plasmodium species analysis. Cytometry. 1987 Nov;8(6):568–570. [PubMed]
  • Martin SK, Oduola AM, Milhous WK. Reversal of chloroquine resistance in Plasmodium falciparum by verapamil. Science. 1987 Feb 20;235(4791):899–901. [PubMed]
  • Oduola AM, Milhous WK, Weatherly NF, Bowdre JH, Desjardins RE. Plasmodium falciparum: induction of resistance to mefloquine in cloned strains by continuous drug exposure in vitro. Exp Parasitol. 1988 Dec;67(2):354–360. [PubMed]
  • Ouellette M, Borst P. Drug resistance and P-glycoprotein gene amplification in the protozoan parasite Leishmania. Res Microbiol. 1991 Jul-Aug;142(6):737–746. [PubMed]

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