Variations in the sequence and expression of the Plasmodium falciparum chloroquine resistance transporter (Pfcrt) and their relationship to chloroquine resistance in vitro

被引:79
|
作者
Durrand, V
Berry, A
Sem, R
Glaziou, P
Beaudou, J
Fandeur, T
机构
[1] Inst Pasteur Cambodge, Lab Epidemiol Mol, Phnom Penh, Cambodia
[2] Fac Med, Serv Parasitol, F-31073 Toulouse 7, France
[3] Natl Malaria Ctr, Phnom Penh, Cambodia
关键词
Plasmodium falciparum; Pfcrt gene; expression level; Cambodia; real-time PCR; chloroquine susceptibility;
D O I
10.1016/j.molbiopara.2004.03.016
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Chloroquine has been widely used for malaria treatment and prophylaxis for several decades, but its usefulness has now declined with the emergence of chloroquine resistance. Recent studies showed that the K76T mutation in the PfCRT protein, initially associated to chloroquine-resistant parasites, is sometimes also present in susceptible parasites, suggesting that other factors control the expression of the resistance phenotype. Here, we sought new mutations in the Pfcrt gene and used real-time PCR to investigate variations in the expression level of this gene with respect to the in vitro response to chloroquine. About 40 Cambodian isolates of Plasmodium falciparum were selected on the basis of their response to chloroquine in vitro. The Pfcrt gene was characterised by amplifying and sequencing the full-length cDNA. Twelve point mutations-M74I, N75D/E, K76T, A144F, L I 481,1194T, A220S, Q271E, N326S, T333S, 1356T and R3711-were detected. Mutations identified at positions 144, 148, 194 and 333 had never been described before. These mutations define six distinct haplotypes, distributed heterogeneously throughout Cambodia. Only the mutations at positions 74-76, 220 and 271 were significantly associated with the in vitro response to chloroquine. Three major haplotypes-MNK/A/Q, IDT/S/E and IET/S/E-accounted for all the isolates examined. The MNK/A/Q haplotype corresponded to susceptible isolates whereas parasites with the IDT/S/E haplotype displayed an intermediate response to chloroquine and those with the IET/S/E haplotype displayed the highest IC50 values. Phylogenic analysis suggested that the IDT and IET haplotypes (positions 74-76) arose independently from the wild-type MNK sequence. We found that the expression level of Pfcrt, evaluated by real-time PCR, had no effect on the response of the parasite to the drug in vitro. Similarly, in a CQ-resistant strain short-term cultured in the presence of CQ, no change was observed in the level of transcripts. These results are discussed in light of recent finding suggesting the possible involvement of other transporters in CQ-resistance. (C) 2004 Elsevier B.V. All rights reserved.
引用
收藏
页码:273 / 285
页数:13
相关论文
共 50 条
  • [41] Phosphomimetic substitution at Ser-33 of the chloroquine resistance transporter PfCRT reconstitutes drug responses in Plasmodium falciparum
    Sanchez, Cecilia P.
    Cubel, Sonia Moliner
    Nyboer, Britta
    Jankowska-Doellken, Monika
    Schaeffer-Reiss, Christine
    Ayoub, Daniel
    Planelles, Gabrielle
    Lanzer, Michael
    [J]. JOURNAL OF BIOLOGICAL CHEMISTRY, 2019, 294 (34) : 12766 - 12778
  • [42] pfcrt polymorphism and the spread of chloroquine resistance in Plasmodium falciparum populations across the Amazon Basin
    Vieira, PP
    Ferreira, MU
    Alecrim, MDGA
    Alecrim, WD
    da Silva, LHP
    Sihuincha, MSM
    Joy, DA
    Mu, JB
    Su, XZ
    Zalis, MG
    [J]. JOURNAL OF INFECTIOUS DISEASES, 2004, 190 (02): : 417 - 424
  • [43] Widespread occurrence of the Plasmodium falciparum chloroquine resistance transporter (PFCRT) gene haplotype SVMNT in P-falciparum malaria in India
    Vathsala, PG
    Pramanik, A
    Dhanasekaran, S
    Devi, CU
    Pillai, CR
    Subbarao, SK
    Ghosh, SK
    Tiwari, SN
    Sathyanarayan, TS
    Deshpande, PR
    Mishra, GC
    Ranjit, MR
    Dash, AP
    Rangarajan, PN
    Padmanaban, G
    [J]. AMERICAN JOURNAL OF TROPICAL MEDICINE AND HYGIENE, 2004, 70 (03): : 256 - 259
  • [44] In vitro and in vivo reversal of chloroquine resistance in Plasmodium falciparum with promethazine
    Oduola, AMJ
    Sowunmi, A
    Milhous, WK
    Brewer, TG
    Kyle, DE
    Gerena, L
    Rossan, RN
    Salako, LA
    Schuster, BG
    [J]. AMERICAN JOURNAL OF TROPICAL MEDICINE AND HYGIENE, 1998, 58 (05): : 625 - 629
  • [45] pfcrt is more than the Plasmodium falciparum chloroquine resistance gene:: a functional and evolutionary perspective
    Cooper, RA
    Hartwig, CL
    Ferdig, MT
    [J]. ACTA TROPICA, 2005, 94 (03) : 170 - 180
  • [46] Is PfCRT a channel or a carrier?: Two competing models explaining chloroquine resistance in Plasmodium falciparum
    Sanchez, Cecilia P.
    Stein, Wilfred D.
    Lanzer, Michael
    [J]. TRENDS IN PARASITOLOGY, 2007, 23 (07) : 332 - 339
  • [47] In vitro reversal of chloroquine resistance in Plasmodium falciparum with dihydroethanoanthracene derivatives
    Pradines, B
    Alibert-Franco, S
    Houdoin, C
    Mosnier, J
    Santelli-Rouvier, C
    Papa, V
    Rogier, C
    Fusai, T
    Barbe, J
    Parzy, D
    [J]. AMERICAN JOURNAL OF TROPICAL MEDICINE AND HYGIENE, 2002, 66 (06): : 661 - 666
  • [48] Mutations in the Plasmodium falciparum chloroquine resistance transporter, PfCRT, enlarge the parasite’s food vacuole and alter drug sensitivities
    Serena Pulcini
    Henry M. Staines
    Andrew H. Lee
    Sarah H. Shafik
    Guillaume Bouyer
    Catherine M. Moore
    Daniel A. Daley
    Matthew J. Hoke
    Lindsey M. Altenhofen
    Heather J. Painter
    Jianbing Mu
    David J. P. Ferguson
    Manuel Llinás
    Rowena E. Martin
    David A. Fidock
    Roland A. Cooper
    Sanjeev Krishna
    [J]. Scientific Reports, 5
  • [49] Mutations in the Plasmodium falciparum chloroquine resistance transporter, PfCRT, enlarge the parasite's food vacuole and alter drug sensitivities
    Pulcini, Serena
    Staines, Henry M.
    Lee, Andrew H.
    Shafik, Sarah H.
    Bouyer, Guillaume
    Moore, Catherine M.
    Daley, Daniel A.
    Hoke, Matthew J.
    Altenhofen, Lindsey M.
    Painter, Heather J.
    Mu, Jianbing
    Ferguson, David J. P.
    Llinas, Manuel
    Martin, Rowena E.
    Fidock, David A.
    Cooper, Roland A.
    Krishna, Sanjeev
    [J]. SCIENTIFIC REPORTS, 2015, 5
  • [50] Prevalence of Plasmodium falciparum pfcrt polymorphisms and in vitro chloroquine sensitivity in Senegal
    Daily, JP
    Roberts, C
    Thomas, SM
    Ndir, O
    Dieng, T
    Mboup, S
    Wirth, DF
    [J]. PARASITOLOGY, 2003, 126 : 401 - 405