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Genetic mapping of targets mediating differential chemical phenotypes in Plasmodium falciparum

Nature Chemical Biology volume 5, pages 765–771 (2009)Cite this article

Abstract

Studies of gene function and molecular mechanisms in Plasmodium falciparum are hampered by difficulties in characterizing and measuring phenotypic differences between individual parasites. We screened seven parasite lines for differences in responses to 1,279 bioactive chemicals. Hundreds of compounds were active in inhibiting parasite growth; 607 differential chemical phenotypes, defined as pairwise IC50 differences of fivefold or more between parasite lines, were cataloged. We mapped major determinants for three differential chemical phenotypes between the parents of a genetic cross, and we identified target genes by fine mapping and testing the responses of parasites in which candidate genes were genetically replaced with mutant alleles. Differential sensitivity to dihydroergotamine methanesulfonate (1), a serotonin receptor antagonist, was mapped to a gene encoding the homolog of human P-glycoprotein (PfPgh-1). This study identifies new leads for antimalarial drugs and demonstrates the utility of a high-throughput chemical genomic strategy for studying malaria traits.

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Figure 1: A chemical genomic strategy for studying gene function in malaria parasites.
Figure 2: Hierarchical clustering of compound activities in seven P. falciparum lines.
Figure 3: Mapping genetic loci contributing to IC50 differences between 7G8 and GB4 in response to dihydroergotamine methanesulfonate (DHMS, 1).
Figure 4: Identification of genetic loci linked to response to trimethoprim (11).

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Acknowledgements

This work was supported by the Divisions of Intramural Research at the National Institute of Allergy and Infectious Diseases, and by the National Human Genome Research Institute and the NIH Roadmap for Medical Research, all at the US National Institutes of Health. We thank J. Sa, J. Mu, L. Jiang, D. Raj and M.J. López Barragán for help in drug assays; P. Shinn and D. VanLeer for compound management; D. Leja for assistance in artworks and NIAID intramural editor B.R. Marshall for assistance.

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Authors and Affiliations

  1. Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases, Bethesda, Maryland, USA

    Jing Yuan, Hongying Jiang, Karen Hayton, Thomas E Wellems & Xin-zhuan Su

  2. NIH Chemical Genomics Center, National Human Genome Research Institute, National Institutes of Health, Bethesda, Maryland, USA

    Ronald L Johnson, Ruili Huang, Jennifer Wichterman, James Inglese & Christopher P Austin

  3. Department of Microbiology and Immunology, Columbia University, New York, New York, USA

    David A Fidock

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Contributions

J.Y. performed drug assay qHTS, parasite culture and data analysis; R.L.J. performed assay optimization, qHTS and data analysis and helped write the manuscript; J.W. performed qHTS; R.H. performed data analysis; H.J. performed drug assays; K.H. performed progeny cloning and helped write the manuscript; D.A.F. transfected parasites and helped write the manuscript; T.E.W. produced progeny and helped write the manuscript; C.P.A. and J.I. planned the project and helped write the manuscript; X.-z.S. conceived the project, analyzed data and helped write the manuscript.

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Correspondence to Xin-zhuan Su.

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Yuan, J., Johnson, R., Huang, R. et al. Genetic mapping of targets mediating differential chemical phenotypes in Plasmodium falciparum. Nat Chem Biol 5, 765–771 (2009). https://doi.org/10.1038/nchembio.215

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