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

Research output: Contribution to journalArticle

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Abstract

Mutations in the Plasmodium falciparum chloroquine resistance transporter, PfCRT, are the major determinant of chloroquine resistance in this lethal human malaria parasite. Here, we describe P. falciparum lines subjected to selection by amantadine or blasticidin that carry PfCRT mutations (C101F or L272F), causing the development of enlarged food vacuoles. These parasites also have increased sensitivity to chloroquine and some other quinoline antimalarials, but exhibit no or minimal change in sensitivity to artemisinins, when compared with parental strains. A transgenic parasite line expressing the L272F variant of PfCRT confirmed this increased chloroquine sensitivity and enlarged food vacuole phenotype. Furthermore, the introduction of the C101F or L272F mutation into a chloroquine-resistant variant of PfCRT reduced the ability of this protein to transport chloroquine by approximately 93 and 82%, respectively, when expressed in Xenopus oocytes. These data provide, at least in part, a mechanistic explanation for the increased sensitivity of the mutant parasite lines to chloroquine. Taken together, these findings provide new insights into PfCRT function and PfCRT-mediated drug resistance, as well as the food vacuole, which is an important target of many antimalarial drugs.

Original languageEnglish (US)
Article number14552
JournalScientific reports
Volume5
DOIs
StatePublished - Sep 30 2015

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Chloroquine
Vacuoles
Parasites
Food
Mutation
Pharmaceutical Preparations
Antimalarials
Artemisinins
Amantadine
Protein Transport
Plasmodium falciparum
Xenopus
Drug Resistance
Malaria
Oocytes
Plasmodium falciparum PfCRT protein
Phenotype

All Science Journal Classification (ASJC) codes

  • General

Cite this

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. ; Llinás, Manuel ; Martin, Rowena E. ; Fidock, David A. ; Cooper, Roland A. ; Krishna, Sanjeev. / Mutations in the Plasmodium falciparum chloroquine resistance transporter, PfCRT, enlarge the parasite's food vacuole and alter drug sensitivities. In: Scientific reports. 2015 ; Vol. 5.
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abstract = "Mutations in the Plasmodium falciparum chloroquine resistance transporter, PfCRT, are the major determinant of chloroquine resistance in this lethal human malaria parasite. Here, we describe P. falciparum lines subjected to selection by amantadine or blasticidin that carry PfCRT mutations (C101F or L272F), causing the development of enlarged food vacuoles. These parasites also have increased sensitivity to chloroquine and some other quinoline antimalarials, but exhibit no or minimal change in sensitivity to artemisinins, when compared with parental strains. A transgenic parasite line expressing the L272F variant of PfCRT confirmed this increased chloroquine sensitivity and enlarged food vacuole phenotype. Furthermore, the introduction of the C101F or L272F mutation into a chloroquine-resistant variant of PfCRT reduced the ability of this protein to transport chloroquine by approximately 93 and 82{\%}, respectively, when expressed in Xenopus oocytes. These data provide, at least in part, a mechanistic explanation for the increased sensitivity of the mutant parasite lines to chloroquine. Taken together, these findings provide new insights into PfCRT function and PfCRT-mediated drug resistance, as well as the food vacuole, which is an important target of many antimalarial drugs.",
author = "Serena Pulcini and Staines, {Henry M.} and Lee, {Andrew H.} and Shafik, {Sarah H.} and Guillaume Bouyer and Moore, {Catherine M.} and Daley, {Daniel A.} and Hoke, {Matthew J.} and Altenhofen, {Lindsey M.} and Painter, {Heather J.} and Jianbing Mu and Ferguson, {David J.P.} and Manuel Llin{\'a}s and Martin, {Rowena E.} and Fidock, {David A.} and Cooper, {Roland A.} and Sanjeev Krishna",
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Pulcini, S, Staines, HM, Lee, AH, Shafik, SH, Bouyer, G, Moore, CM, Daley, DA, Hoke, MJ, Altenhofen, LM, Painter, HJ, Mu, J, Ferguson, DJP, Llinás, M, Martin, RE, Fidock, DA, Cooper, RA & Krishna, S 2015, 'Mutations in the Plasmodium falciparum chloroquine resistance transporter, PfCRT, enlarge the parasite's food vacuole and alter drug sensitivities', Scientific reports, vol. 5, 14552. https://doi.org/10.1038/srep14552

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.; Llinás, Manuel; Martin, Rowena E.; Fidock, David A.; Cooper, Roland A.; Krishna, Sanjeev.

In: Scientific reports, Vol. 5, 14552, 30.09.2015.

Research output: Contribution to journalArticle

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T1 - Mutations in the Plasmodium falciparum chloroquine resistance transporter, PfCRT, enlarge the parasite's food vacuole and alter drug sensitivities

AU - Pulcini, Serena

AU - Staines, Henry M.

AU - Lee, Andrew H.

AU - Shafik, Sarah H.

AU - Bouyer, Guillaume

AU - Moore, Catherine M.

AU - Daley, Daniel A.

AU - Hoke, Matthew J.

AU - Altenhofen, Lindsey M.

AU - Painter, Heather J.

AU - Mu, Jianbing

AU - Ferguson, David J.P.

AU - Llinás, Manuel

AU - Martin, Rowena E.

AU - Fidock, David A.

AU - Cooper, Roland A.

AU - Krishna, Sanjeev

PY - 2015/9/30

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N2 - Mutations in the Plasmodium falciparum chloroquine resistance transporter, PfCRT, are the major determinant of chloroquine resistance in this lethal human malaria parasite. Here, we describe P. falciparum lines subjected to selection by amantadine or blasticidin that carry PfCRT mutations (C101F or L272F), causing the development of enlarged food vacuoles. These parasites also have increased sensitivity to chloroquine and some other quinoline antimalarials, but exhibit no or minimal change in sensitivity to artemisinins, when compared with parental strains. A transgenic parasite line expressing the L272F variant of PfCRT confirmed this increased chloroquine sensitivity and enlarged food vacuole phenotype. Furthermore, the introduction of the C101F or L272F mutation into a chloroquine-resistant variant of PfCRT reduced the ability of this protein to transport chloroquine by approximately 93 and 82%, respectively, when expressed in Xenopus oocytes. These data provide, at least in part, a mechanistic explanation for the increased sensitivity of the mutant parasite lines to chloroquine. Taken together, these findings provide new insights into PfCRT function and PfCRT-mediated drug resistance, as well as the food vacuole, which is an important target of many antimalarial drugs.

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