Chloroquine resistance modulated in vitro by expression levels of the Plasmodium falciparum chloroquine resistance transporter - PubMed (original) (raw)
. 2003 Aug 29;278(35):33593-601.
doi: 10.1074/jbc.M302215200. Epub 2003 Jun 17.
Affiliations
- PMID: 12813054
- DOI: 10.1074/jbc.M302215200
Free article
Chloroquine resistance modulated in vitro by expression levels of the Plasmodium falciparum chloroquine resistance transporter
Karena L Waller et al. J Biol Chem. 2003.
Free article
Abstract
Plasmodium falciparum malaria is increasingly difficult to treat and control due to the emergence of parasite resistance to the major antimalarials, notably chloroquine. Recent work has shown that the chloroquine resistance phenotype can be conferred by multiple amino acid mutations in the parasite digestive vacuole transmembrane protein PfCRT. Here, we have addressed whether chloroquine resistance can also be affected by changes in expression levels of this protein. Transient transfection reporter assays revealed that truncation of the pfcrt 3'-untranslated region just prior to putative polyadenylation sites resulted in a 10-fold decrease in luciferase expression levels. Using allelic exchange on a chloroquine-resistant line (7G8 from Brazil), this truncated 3'-untranslated region was inserted downstream of the pfcrt coding sequence, in the place of the endogenous 3'-untranslated region. The resulting pfcrt-modified "knockdown" clones displayed a marked decrease in pfcrt transcription and an estimated 30-40% decrease in PfCRT protein expression levels. [3H]hypoxanthine incorporation assays demonstrated up to a 40% decrease in chloroquine with or without verapamil IC50 levels of pfcrt knockdown clones, relative to the 7G8 parent. Single-cell photometric analyses were consistent with an altered intracellular pH in the knockdown clones, providing further evidence for a relationship between PfCRT, pH regulation, and chloroquine resistance. Genetic truncation of 3'-untranslated regions provides a useful approach for assessing the impact of candidate genes on drug resistance or other quantifiable phenotypes in P. falciparum.
Similar articles
- Combinatorial Genetic Modeling of pfcrt-Mediated Drug Resistance Evolution in Plasmodium falciparum.
Gabryszewski SJ, Modchang C, Musset L, Chookajorn T, Fidock DA. Gabryszewski SJ, et al. Mol Biol Evol. 2016 Jun;33(6):1554-70. doi: 10.1093/molbev/msw037. Epub 2016 Feb 22. Mol Biol Evol. 2016. PMID: 26908582 Free PMC article. - Defining the role of PfCRT in Plasmodium falciparum chloroquine resistance.
Bray PG, Martin RE, Tilley L, Ward SA, Kirk K, Fidock DA. Bray PG, et al. Mol Microbiol. 2005 Apr;56(2):323-33. doi: 10.1111/j.1365-2958.2005.04556.x. Mol Microbiol. 2005. PMID: 15813727 Review. - Chloroquine-resistant isoforms of the Plasmodium falciparum chloroquine resistance transporter acidify lysosomal pH in HEK293 cells more than chloroquine-sensitive isoforms.
Reeves DC, Liebelt DA, Lakshmanan V, Roepe PD, Fidock DA, Akabas MH. Reeves DC, et al. Mol Biochem Parasitol. 2006 Dec;150(2):288-99. doi: 10.1016/j.molbiopara.2006.09.001. Epub 2006 Sep 25. Mol Biochem Parasitol. 2006. PMID: 17014918 Free PMC article. - A 2-amino quinoline, 5-(3-(2-(7-chloroquinolin-2-yl)ethenyl)phenyl)-8-dimethylcarbamyl-4,6-dithiaoctanoic acid, interacts with PfMDR1 and inhibits its drug transport in Plasmodium falciparum.
Edaye S, Reiling SJ, Leimanis ML, Wunderlich J, Rohrbach P, Georges E. Edaye S, et al. Mol Biochem Parasitol. 2014 Jun;195(1):34-42. doi: 10.1016/j.molbiopara.2014.05.006. Epub 2014 Jun 8. Mol Biochem Parasitol. 2014. PMID: 24914817 - Is PfCRT a channel or a carrier? Two competing models explaining chloroquine resistance in Plasmodium falciparum.
Sanchez CP, Stein WD, Lanzer M. Sanchez CP, et al. Trends Parasitol. 2007 Jul;23(7):332-9. doi: 10.1016/j.pt.2007.04.013. Epub 2007 May 10. Trends Parasitol. 2007. PMID: 17493873 Review.
Cited by
- Design, synthesis, and evaluation of 10-N-substituted acridones as novel chemosensitizers in Plasmodium falciparum.
Kelly JX, Smilkstein MJ, Cooper RA, Lane KD, Johnson RA, Janowsky A, Dodean RA, Hinrichs DJ, Winter R, Riscoe M. Kelly JX, et al. Antimicrob Agents Chemother. 2007 Nov;51(11):4133-40. doi: 10.1128/AAC.00669-07. Epub 2007 Sep 10. Antimicrob Agents Chemother. 2007. PMID: 17846138 Free PMC article. - Targeting protein translation, RNA splicing, and degradation by morpholino-based conjugates in Plasmodium falciparum.
Garg A, Wesolowski D, Alonso D, Deitsch KW, Ben Mamoun C, Altman S. Garg A, et al. Proc Natl Acad Sci U S A. 2015 Sep 22;112(38):11935-40. doi: 10.1073/pnas.1515864112. Epub 2015 Sep 8. Proc Natl Acad Sci U S A. 2015. PMID: 26351679 Free PMC article. - Therapeutic efficacy of artemether-lumefantrine for the treatment of uncomplicated Plasmodium falciparum malaria in Enfranze, north-west Ethiopia.
Getnet G, Fola AA, Alemu A, Getie S, Fuehrer HP, Noedl H. Getnet G, et al. Malar J. 2015 Jun 24;14:258. doi: 10.1186/s12936-015-0775-3. Malar J. 2015. PMID: 26105035 Free PMC article. - The Knock-Down of the Chloroquine Resistance Transporter PfCRT Is Linked to Oligopeptide Handling in Plasmodium falciparum.
Sanchez CP, Manson EDT, Moliner Cubel S, Mandel L, Weidt SK, Barrett MP, Lanzer M. Sanchez CP, et al. Microbiol Spectr. 2022 Aug 31;10(4):e0110122. doi: 10.1128/spectrum.01101-22. Epub 2022 Jul 18. Microbiol Spectr. 2022. PMID: 35867395 Free PMC article. - Identification of the drug/metabolite transporter 1 as a marker of quinine resistance in a NF54×Cam3.II P. falciparum genetic cross.
Kanai M, Mok S, Yeo T, Shears MJ, Ross LS, Jeon JH, Narwal S, Haile MT, Tripathi AK, Mlambo G, Kim J, Gil-Iturbe E, Okombo J, Fairhurst KJ, Bloxham T, Bridgford JL, Sheth T, Ward KE, Park H, Rozenberg FD, Quick M, Mancia F, Lee MCS, Small-Saunders JL, Uhlemann AC, Sinnis P, Fidock DA. Kanai M, et al. bioRxiv [Preprint]. 2024 Oct 1:2024.09.27.615529. doi: 10.1101/2024.09.27.615529. bioRxiv. 2024. PMID: 39386571 Free PMC article. Preprint.
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources