Phenotypic and transcriptomic analyses of Plasmodium falciparum protein kinase A catalytic subunit inhibition.
The emergence and dissemination of drug-resistant malaria parasites represent one of the most important problems in malaria case management. Plasmodium falciparum is the causative agent of the most lethal form of human malaria. The molecular mechanisms that control the life cycle of the malaria parasite are still poorly understood. The published genome sequence (P. falciparum strain 3D7) reveals that several homologs of eukaryotic signaling proteins, such as protein kinases and phosphatases, are conserved in P. falciparum. Proteins kinases are now widely recognized as valuable drug targets in protozoan parasites. In this study, gene silencing with double-stranded RNA (dsRNA) and microarray techniques were used to study the biological function of the cAMP-dependent protein kinase catalytic subunit (PfPKAc) in the parasite erythrocytic life cycle. Treatment of parasites with PfPKAc dsRNA resulted in a marked reduction of endogenous PfPKAc mRNA associated with a compensatory decrease of PfPKAr mRNA followed by morphological changes in schizont stages and cell cycle arrest. The global effects of gene silencing were also investigated using a P. falciparum pan-genomic microarray. Transcriptomic analysis showed that the expression of 329 genes was altered in response to downregulation of PfPKAc mRNA particularly genes in specific metabolic pathways linked with merozoite invasion processes, the calcium/calmodulin signaling, and kinases network and mitochondrial functions.
UMR-MD3, Relation Hôte-Parasites-Pharmacologie et Thérapeutique (Université de la Méditerranée Aix-Marseille II), des Armées-Antenne Marseille, Institut de Recherche Biomédicale/IMTSSA, Allée du Médecin colonel Eugène Jamot, Parc du Pharo, Marseille, Cedex 07, France.
Nathalie
N
Wurtz
Jérôme
J
Desplans
Daniel
D
Parzy
D000818:Animals; D020134:Catalytic Domain; D002453:Cell Cycle / Q000187:drug effects; D017868:Cyclic AMP-Dependent Protein Kinases / Q000037:antagonists & inhibitors* / Q000235:genetics; D015536:Down-Regulation; D020868:Gene Silencing; D020411:Oligonucleotide Array Sequence Analysis; D010963:Plasmodium falciparum / Q000737:chemistry / Q000166:cytology / Q000201:enzymology* / Q000235:genetics; D012330:RNA, Double-Stranded / Q000235:genetics; D012333:RNA, Messenger / Q000096:biosynthesis; D016053:RNA, Protozoan / Q000096:biosynthesis
D012330:RNA, Double-Stranded; D012333:RNA, Messenger; D016053:RNA, Protozoan; D017868:Cyclic AMP-Dependent Protein Kinases