Evidence for Regulation of Hemoglobin Metabolism and Intracellular Ionic Flux by the Plasmodium falciparum Chloroquine Resistance Transporter

Andrew H. Lee, Satish K. Dhingra, Ian A. Lewis, Maneesh K. Singh, Amila Siriwardana, Seema Dalal, Kelly Rubiano, Matthias S. Klein, Katelynn S. Baska, Sanjeev Krishna, Michael Klemba, Paul D. Roepe, Manuel Llinás, Celia R.S. Garcia, David A. Fidock

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8 Scopus citations

Abstract

Plasmodium falciparum multidrug resistance constitutes a major obstacle to the global malaria elimination campaign. Specific mutations in the Plasmodium falciparum chloroquine resistance transporter (PfCRT) mediate resistance to the 4-aminoquinoline drug chloroquine and impact parasite susceptibility to several partner agents used in current artemisinin-based combination therapies, including amodiaquine. By examining gene-edited parasites, we report that the ability of the wide-spread Dd2 PfCRT isoform to mediate chloroquine and amodiaquine resistance is substantially reduced by the addition of the PfCRT L272F mutation, which arose under blasticidin selection. We also provide evidence that L272F confers a significant fitness cost to asexual blood stage parasites. Studies with amino acid-restricted media identify this mutant as a methionine auxotroph. Metabolomic analysis also reveals an accumulation of short, hemoglobin-derived peptides in the Dd2 + L272F and Dd2 isoforms, compared with parasites expressing wild-type PfCRT. Physiologic studies with the ionophores monensin and nigericin support an impact of PfCRT isoforms on Ca2+ release, with substantially reduced Ca2+ levels observed in Dd2 + L272F parasites. Our data reveal a central role for PfCRT in regulating hemoglobin catabolism, amino acid availability, and ionic balance in P. falciparum, in addition to its role in determining parasite susceptibility to heme-binding 4-aminoquinoline drugs.

Original languageEnglish (US)
Article number13578
JournalScientific reports
Volume8
Issue number1
DOIs
StatePublished - Dec 1 2018

All Science Journal Classification (ASJC) codes

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    Lee, A. H., Dhingra, S. K., Lewis, I. A., Singh, M. K., Siriwardana, A., Dalal, S., Rubiano, K., Klein, M. S., Baska, K. S., Krishna, S., Klemba, M., Roepe, P. D., Llinás, M., Garcia, C. R. S., & Fidock, D. A. (2018). Evidence for Regulation of Hemoglobin Metabolism and Intracellular Ionic Flux by the Plasmodium falciparum Chloroquine Resistance Transporter. Scientific reports, 8(1), [13578]. https://doi.org/10.1038/s41598-018-31715-9