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.

Details

Title
Evidence for Regulation of Hemoglobin Metabolism and Intracellular Ionic Flux by the Plasmodium falciparum Chloroquine Resistance Transporter
Author
Lee, Andrew H 1 ; Dhingra, Satish K 1 ; Lewis, Ian A 2 ; Singh, Maneesh K 3 ; Siriwardana, Amila 4 ; Dalal, Seema 5 ; Rubiano, Kelly 1 ; Klein, Matthias S 6 ; Baska, Katelynn S 7 ; Krishna, Sanjeev 8   VIAFID ORCID Logo  ; Klemba, Michael 5 ; Roepe, Paul D 4 ; Llinás, Manuel 9 ; Garcia, Celia R S 10 ; Fidock, David A 11 

1  Department of Microbiology and Immunology, Columbia University Medical Center, New York, NY, USA 
2  Department of Biological Sciences, University of Calgary, Calgary, Canada; Lewis-Sigler Institute for Integrative Genomics, Princeton University, Princeton, NJ, USA 
3  Department of Parasitology, Institute of Biomedical Sciences, University of São Paulo, São Paulo, Brazil; Department of Physiology and Biophysics, Institute of Biomedical Sciences, University of São Paulo, São Paulo, Brazil 
4  Departments of Chemistry and of Biochemistry and Cellular & Molecular Biology, Georgetown University, Washington, DC, USA 
5  Department of Biochemistry, Virginia Tech, Blacksburg, VA, USA 
6  Department of Biological Sciences, University of Calgary, Calgary, Canada 
7  Lewis-Sigler Institute for Integrative Genomics, Princeton University, Princeton, NJ, USA 
8  Institute for Infection and Immunity, St. George’s University Hospital, University of London, London, UK 
9  Departments of Chemistry and of Biochemistry and Molecular Biology and Center for Malaria Research, Pennsylvania State University, State College, PA, USA 
10  Department of Physiology and Biophysics, Institute of Biomedical Sciences, University of São Paulo, São Paulo, Brazil; Department of Clinical and Toxicological Analyses, School of Pharmaceutical Sciences, University of São Paulo, São Paulo, Brazil 
11  Department of Microbiology and Immunology, Columbia University Medical Center, New York, NY, USA; Division of Infectious Diseases, Department of Medicine, Columbia University Medical Center, New York, NY, USA 
Pages
1-13
Publication year
2018
Publication date
Sep 2018
Publisher
Nature Publishing Group
e-ISSN
20452322
Source type
Scholarly Journal
Language of publication
English
DOI
https://doi.org/10.1038/s41598-018-31715-9
ProQuest document ID
2102373994
Copyright
© 2018. This work is published under http://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.