Essentiality of Plasmodium falciparum plasmepsin V

PLoS One. 2018 Dec 5;13(12):e0207621. doi: 10.1371/journal.pone.0207621. eCollection 2018.

Abstract

The malaria parasite replicates within erythrocytes. The pathogenesis of clinical malaria is in large part due to the capacity of the parasite to remodel its host cell. To do this, intraerythrocytic stages of Plasmodium falciparum export more than 300 proteins that dramatically alter the morphology of the infected erythrocyte as well as its mechanical and adhesive properties. P. falciparum plasmepsin V (PfPMV) is an aspartic protease that processes proteins for export into the host erythrocyte and is thought to play a key role in parasite virulence and survival. However, although standard techniques for gene disruption as well as conditional protein knockdown have been previously attempted with the pfpmv gene, complete gene removal or knockdown was not achieved so direct genetic proof that PMV is an essential protein has not been established. Here we have used a conditional gene excision approach combining CRISPR-Cas9 gene editing and DiCre-mediated recombination to functionally inactivate the pfpmv gene. The resulting mutant parasites displayed a severe growth defect. Detailed phenotypic analysis showed that development of the mutant parasites was arrested early in the ring-to-trophozoite transition in the erythrocytic cycle following gene excision. Our findings are the first to elucidate the effects of PMV gene disruption, showing that it is essential for parasite viability in asexual blood stages. The mutant parasites can now be used as a platform to further dissect the Plasmodium protein export pathway.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aspartic Acid Endopeptidases / genetics*
  • Aspartic Acid Endopeptidases / metabolism
  • Aspartic Acid Endopeptidases / physiology*
  • CRISPR-Cas Systems
  • Erythrocytes / metabolism
  • Erythrocytes / parasitology
  • Gene Expression Regulation
  • Humans
  • Mutation / genetics
  • Plasmodium falciparum / genetics
  • Plasmodium falciparum / metabolism*
  • Protease Inhibitors
  • Protein Processing, Post-Translational
  • Protozoan Proteins / metabolism

Substances

  • Protease Inhibitors
  • Protozoan Proteins
  • Aspartic Acid Endopeptidases
  • plasmepsin