1887

Abstract

The kinome includes a family of four protein kinases (Pfnek-1 to -4) related to the NIMA (never-in-mitosis) family, members of which play important roles in mitosis and meiosis in eukaryotic cells. Only one of these, Pfnek-1, which we previously characterized at the biochemical level, is expressed in asexual parasites. The other three (Pfnek-2, -3 and -4) are expressed predominantly in gametocytes, and a role for nek-2 and nek-4 in meiosis has been documented. Here we show by reverse genetics that Pfnek-1 is required for completion of the asexual cycle in red blood cells and that its expression in gametocytes in detectable by immunofluorescence in male (but not in female) gametocytes, in contrast with Pfnek-2 and Pfnek-4. This indicates that the function of Pfnek-1 is non-redundant with those of the other members of the Pfnek family and identifies Pfnek-1 as a potential target for antimalarial chemotherapy. A medium-throughput screen of a small-molecule library provides proof of concept that recombinant Pfnek-1 can be used as a target in drug discovery.

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2011-10-01
2019-09-21
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References

  1. Abdi A. , Eschenlauer S. , Reininger L. , Doerig C. . ( 2010; ). SAM domain-dependent activity of PfTKL3, an essential tyrosine kinase-like kinase of the human malaria parasite Plasmodium falciparum . . Cell Mol Life Sci 67:, 3355–3369. [CrossRef] [PubMed]
    [Google Scholar]
  2. Alano P , Read D. , Bruce M. , Aikawa M. , Kaido T. , Tegoshi T. , Bhatti S. , Smith D. K. , Luo C. et al. ( 1995; ). COS cell expression cloning of Pfg377, a Plasmodium falciparum gametocyte antigen associated with osmiophilic bodies. . Mol Biochem Parasitol 74:, 143–156. [CrossRef] [PubMed]
    [Google Scholar]
  3. Anamika S. , Srinivasan N , Krupa A. . ( 2005; ). A genomic perspective of protein kinases in Plasmodium falciparum . . Proteins 58:, 180–189. [CrossRef] [PubMed]
    [Google Scholar]
  4. Carter R. , Ranford-Cartwright L. , Alano P . ( 1993; ). The culture and preparation of gametocytes of Plasmodium falciparum for immunochemical, molecular, and mosquito infectivity studies. . Methods Mol Biol 21:, 67–88.[PubMed]
    [Google Scholar]
  5. Carvalho T. G. , Ménard R. . ( 2005; ). Manipulating the Plasmodium genome. . Curr Issues Mol Biol 7:, 39–55.[PubMed]
    [Google Scholar]
  6. de Koning-Ward T. F. , Olivieri A. , Bertuccini L. , Hood A. , Silvestrini F. , Charvalias K. , Berzosa Díaz P. , Camarda G. , McElwain T. F. et al. ( 2008; ). The role of osmiophilic bodies and Pfg377 expression in female gametocyte emergence and mosquito infectivity in the human malaria parasite Plasmodium falciparum . . Mol Microbiol 67:, 278–290. [CrossRef] [PubMed]
    [Google Scholar]
  7. Desoubzdanne D. , Marcourt L. , Raux R. , Chevalley S. , Dorin D. , Doerig C. , Valentin A. , Ausseil F. , Debitus C. . ( 2008; ). Alisiaquinones and alisiaquinol, dual inhibitors of Plasmodium falciparum enzyme targets from a New Caledonian deep water sponge. . J Nat Prod 71:, 1189–1192. [CrossRef] [PubMed]
    [Google Scholar]
  8. Doerig C. , Abdi A. , Bland N. , Eschenlauer S. , Dorin-Semblat D. , Fennell C. , Halbert J. , Holland Z. , Nivez M. P. et al. ( 2010; ). Malaria: targeting parasite and host cell kinomes. . Biochim Biophys Acta 1804:, 604–612.[PubMed] [CrossRef]
    [Google Scholar]
  9. Dorin D , Le Roch K. , Sallicandro P. , Alano P. , Parzy D. , Poullet P. , Meijer L. , Doerig C. . ( 2001; ). Pfnek-1, a NIMA-related kinase from the human malaria parasite Plasmodium falciparum. Biochemical properties and possible involvement in MAPK regulation. . Eur J Biochem 268:, 2600–2608. [CrossRef] [PubMed]
    [Google Scholar]
  10. Dorin D , Semblat J. P. , Poullet P. , Alano P. , Goldring J. P. , Whittle C. , Patterson S. , Chakrabarti D. , Doerig C. . ( 2005; ). PfPK7, an atypical MEK-related protein kinase, reflects the absence of classical three-component MAPK pathways in the human malaria parasite Plasmodium falciparum . . Mol Microbiol 55:, 184–186. [CrossRef] [PubMed]
    [Google Scholar]
  11. Dorin-Semblat D. , Quashie N. , Halbert J. , Sicard A. , Doerig C. , Peat E. , Ranford-Cartwright L. , Doerig C. . ( 2007; ). Functional characterization of both MAP kinases of the human malaria parasite Plasmodium falciparum by reverse genetics. . Mol Microbiol 65:, 1170–1180. [CrossRef] [PubMed]
    [Google Scholar]
  12. Dorin-Semblat D. , Sicard A. , Doerig C. , Ranford-Cartwright L. , Doerig C. . ( 2008; ). Disruption of the PfPK7 gene impairs schizogony and sporogony in the human malaria parasite Plasmodium falciparum . . Eukaryot Cell 7:, 279–285. [CrossRef] [PubMed]
    [Google Scholar]
  13. Dvorin J. D. , Martyn D. C. , Patel S. D. , Grimley J. S. , Collins C. R. , Hopp C. S. , Bright A. T. , Westenberger S. , Winzeler E. et al. ( 2010; ). A plant-like kinase in Plasmodium falciparum regulates parasite egress from erythrocytes. . Science 328:, 910–912. [CrossRef] [PubMed]
    [Google Scholar]
  14. Fennell C. , Babbitt S. , Russo I. , Wilkes J. , Ranford-Cartwright L. , Goldberg D. E. , Doerig C. . ( 2009; ). PfeIK1, a eukaryotic initiation factor 2α kinase of the human malaria parasite Plasmodium falciparum, regulates stress-response to amino-acid starvation. . Malar J 8:, 99. [CrossRef] [PubMed]
    [Google Scholar]
  15. Fry A. M. , Meraldi P. , Nigg E. A. . ( 1998; ). A centrosomal function for the human Nek2 protein kinase, a member of the NIMA family of cell cycle regulators. . EMBO J 17:, 470–481. [CrossRef] [PubMed]
    [Google Scholar]
  16. Gamo F. J. , Sanz L. M. , Vidal J. , de Cozar C. , Alvarez E. , Lavandera J. L. , Vanderwall D. E. , Green D. V. , Kumar V. et al. ( 2010; ). Thousands of chemical starting points for antimalarial lead identification. . Nature 465:, 305–310. [CrossRef] [PubMed]
    [Google Scholar]
  17. Guiguemde W. A. , Shelat A. A. , Bouck D. , Duffy S. , Crowther G. J. , Davis P. H. , Smithson D. C. , Connelly M. , Clark J. et al. ( 2010; ). Chemical genetics of Plasmodium falciparum . . Nature 465:, 311–315. [CrossRef] [PubMed]
    [Google Scholar]
  18. Halbert J. , Ayong L. , Equinet L. , Le Roch K. , Hardy M. , Goldring D. , Reininger L. , Waters N. , Chakrabarti D. , Doerig C. . ( 2010; ). A Plasmodium falciparum transcriptional cyclin-dependent kinase-related kinase with a crucial role in parasite proliferation associates with histone deacetylase activity. . Eukaryot Cell 9:, 952–959. [CrossRef] [PubMed]
    [Google Scholar]
  19. Holland Z. , Prudent R. , Reiser J. B. , Cochet C. , Doerig C. . ( 2009; ). Functional analysis of protein kinase CK2 of the human malaria parasite Plasmodium falciparum . . Eukaryot Cell 8:, 388–397. [CrossRef] [PubMed]
    [Google Scholar]
  20. Khan S. M. , Franke-Fayard B. , Mair G. R. , Lasonder E. , Janse C. J. , Mann M. , Waters A. P. . ( 2005; ). Proteome analysis of separated male and female gametocytes reveals novel sex-specific Plasmodium biology. . Cell 121:, 675–687. [CrossRef] [PubMed]
    [Google Scholar]
  21. Kissinger J. C. , Brunk B. P. , Crabtree J. , Fraunholz M. J. , Gajria B. , Milgram A. J. , Pearson D. S. , Schug J. , Bahl A. et al. ( 2002; ). The Plasmodium genome database. . Nature 419:, 490–492. [CrossRef] [PubMed]
    [Google Scholar]
  22. Lambros C. , Vanderberg J P. . ( 1979; ). Synchronization of Plasmodium falciparum erythrocytic stages in culture. . J Parasitol 65:, 418–420. [CrossRef] [PubMed]
    [Google Scholar]
  23. Laurent D. , Jullian V. , Parenty A. , Knibiehler M. , Dorin D. , Schmitt S. , Lozach O. , Lebouvier N. , Frostin M. , Alby F. . ( 2006; ). Antimalarial potential of xestoquinone, a protein kinase inhibitor isolated from a Vanuatu marine sponge Xestospongia sp. . Bioorg Med Chem 14:, 4477–4482. [CrossRef] [PubMed]
    [Google Scholar]
  24. Le Roch K. G. , Zhou Y. , Blair P. L. , Grainger M. , Moch J. K. , Haynes J. D. , De La Vega P. , Holder A. A. , Batalov S. et al. ( 2003; ). Discovery of gene function by expression profiling of the malaria parasite life cycle. . Science 301:, 1503–1508. [CrossRef] [PubMed]
    [Google Scholar]
  25. Le Roch K. G. , Johnson J. R. , Florens L. , Zhou Y. , Santrosyan A. , Grainger M. , Yan S. F. , Williamson K. C. , Holder A. A. et al. ( 2004; ). Global analysis of transcript and protein levels across the Plasmodium falciparum life cycle. . Genome Res 14:, 2308–2318. [CrossRef] [PubMed]
    [Google Scholar]
  26. Madeira L. , Galante P. A. , Budu A. , Azevedo M. F. , Malnic B. , Garcia C. R. . ( 2008; ). Genome-wide detection of serpentine receptor-like proteins in malaria parasites. . PLoS ONE 3:, e1889. [CrossRef] [PubMed]
    [Google Scholar]
  27. McRobert L. , Taylor C. J. , Deng W. , Fivelman Q. L. , Cummings R. M. , Polley S. D. , Billker O. , Baker D. A. . ( 2008; ). Gametogenesis in malaria parasites is mediated by the cGMP-dependent protein kinase. . PLoS Biol 6:, e139. [CrossRef] [PubMed]
    [Google Scholar]
  28. Merckx A. , Nivez M. P. , Bouyer G. , Alano P. , Langsley G. , Deitsch K. , Thomas S. , Doerig C. , Egée S. . ( 2008; ). Plasmodium falciparum regulatory subunit of cAMP-dependent PKA and anion channel conductance. . PLoS Pathog 4:, e19. [CrossRef] [PubMed]
    [Google Scholar]
  29. O’Regan L. , Blot J , Fry A. M. . ( 2007; ). Mitotic regulation by NIMA-related kinases. . Cell Div 2:, 25. [CrossRef] [PubMed]
    [Google Scholar]
  30. Plouffe D. , Brinker A. , McNamara C. , Henson K. , Kato N. , Kuhen K. , Nagle A. , Adrián F. , Matzen J. T. et al. ( 2008; ). In silico activity profiling reveals the mechanism of action of antimalarials discovered in a high-throughput screen. . Proc Natl Acad Sci U S A 105:, 9059–9064. [CrossRef] [PubMed]
    [Google Scholar]
  31. Rangarajan R. , Bei A. K. , Jethwaney D. , Maldonado P. , Dorin D. , Sultan A. A. , Doerig C. . ( 2005; ). A mitogen-activated protein kinase regulates male gametogenesis and transmission of the malaria parasite Plasmodium berghei . . EMBO Rep 6:, 464–469. [CrossRef] [PubMed]
    [Google Scholar]
  32. Rangarajan R. , Bei A. , Henry N. , Madamet M. , Parzy D. , Nivez M. P. , Doerig C. , Sultan A. . ( 2006; ). Pbcrk-1, the Plasmodium berghei orthologue of P. falciparum cdc-2 related kinase-1 (Pfcrk-1), is essential for completion of the intraerythrocytic asexual cycle. . Exp Parasitol 112:, 202–207. [CrossRef] [PubMed]
    [Google Scholar]
  33. Reininger L. , Billker O. , Tewari R. , Mukhopadhyay A. , Fennell C. , Dorin-Semblat D. , Doerig C. , Goldring D. , Harmse L. et al. ( 2005; ). A NIMA-related protein kinase is essential for completion of the sexual cycle of malaria parasites. . J Biol Chem 280:, 31957–31964. [CrossRef] [PubMed]
    [Google Scholar]
  34. Reininger L. , Tewari R. , Fennell C. , Holland Z. , Goldring D. , Ranford-Cartwright L. , Billker O. , Doerig C. . ( 2009; ). An essential role for the Plasmodium Nek-2 Nima-related protein kinase in the sexual development of malaria parasites. . J Biol Chem 284:, 20858–20868. [CrossRef] [PubMed]
    [Google Scholar]
  35. Rhee K. , Wolgemuth D. J. . ( 1997; ). The NIMA-related kinase 2, Nek2, is expressed in specific stages of the meiotic cell cycle and associates with meiotic chromosomes. . Development 124:, 2167–2177.[PubMed]
    [Google Scholar]
  36. Sathornsumetee S. , Reardon D. A. . ( 2009; ). Targeting multiple kinases in glioblastoma multiforme. . Expert Opin Investig Drugs 18:, 277–292. [CrossRef] [PubMed]
    [Google Scholar]
  37. Schwank S. , Sutherland C J. , Drakeley C. J. . ( 2010; ). Promiscuous expression of α-tubulin II in maturing male and female Plasmodium falciparum gametocytes. . PLoS ONE 5:, e14470. [CrossRef] [PubMed]
    [Google Scholar]
  38. Sidhu A B. , Valderramos S G. , Fidock D. A. . ( 2005; ). pfmdr1 mutations contribute to quinine resistance and enhance mefloquine and artemisinin sensitivity in Plasmodium falciparum. . Mol Microbiol 57:, 913–926. [CrossRef] [PubMed]
    [Google Scholar]
  39. Taylor H. M. , McRobert L. , Grainger M. , Sicard A. , Dluzewski A. R. , Hopp C. S. , Holder A. A. , Baker D. A. . ( 2010; ). The malaria parasite cyclic GMP-dependent protein kinase plays a central role in blood-stage schizogony. . Eukaryot Cell 9:, 37–45. [CrossRef] [PubMed]
    [Google Scholar]
  40. Tewari R. , Dorin D. , Moon R. , Doerig C. , Billker O. . ( 2005; ). An atypical mitogen-activated protein kinase controls cytokinesis and flagellar motility during male gamete formation in a malaria parasite. . Mol Microbiol 58:, 1253–1263. [CrossRef] [PubMed]
    [Google Scholar]
  41. Tewari R. , Straschil U. , Bateman A. , Böhme U. , Cherevach I. , Gong P. , Pain A. , Billker O. . ( 2010; ). The systematic functional analysis of Plasmodium protein kinases identifies essential regulators of mosquito transmission. . Cell Host Microbe 8:, 377–387. [CrossRef] [PubMed]
    [Google Scholar]
  42. Ward P. , Equinet L. , Packer J. , Doerig C. . ( 2004; ). Protein kinases of the human malaria parasite Plasmodium falciparum: the kinome of a divergent eukaryote. . BMC Genomics 5:, 79. [CrossRef] [PubMed]
    [Google Scholar]
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