A divergent cyclin/cyclin-dependent kinase complex controls the atypical replication of a malaria parasite during gametogony and transmission

Abstract

Cell cycle transitions are generally triggered by variation in the activity of cyclin-dependent kinases (CDKs) bound to cyclins. Malaria-causing parasites have a life cycle with unique cell-division cycles, and a repertoire of divergent CDKs and cyclins of poorly understood function and interdependency. We show that Plasmodium berghei CDK-related kinase 5 (CRK5), is a critical regulator of atypical mitosis in the gametogony and is required for mosquito transmission. It phosphorylates canonical CDK motifs of components in the pre-replicative complex and is essential for DNA replication. During a replicative cycle, CRK5 stably interacts with a single Plasmodium-specific cyclin (SOC2), although we obtained no evidence of SOC2 cycling by transcription, translation or degradation. Our results provide evidence that during Plasmodium male gametogony, this divergent cyclin/CDK pair fills the functional space of other eukaryotic cell-cycle kinases controlling DNA replication.

Protein Phosphatase 1 regulates atypical mitotic and meiotic division inPlasmodiumsexual stages

“…flagella formation). A similar phenotype was observed in our recent study of a divergent Plasmodium cdc2-related kinase (CRK5) 54 . CRK5 deletion resulted in fewer nuclear poles, and no chromatin condensation, cytokinesis or flagellum formation, suggesting that there may be a coordinated activity of PP1 and CRK5 in reversible phosphorylation.…”

Expansion microscopy provides new insights into the cytoskeleton of malaria parasites including the conservation of a conoid

“…P . berghei ANKA strain-derived [ 45 ] clone 2.34 [ 46 ] together with derived transgenic lines Kin8B-KO [ 24 ], SAS6L-GFP [ 36 ], and MyoB-GFP [ 37 ] were grown and maintained in CD1 outbred mice as previously described [ 47 ]. Moreover, 6- to 10-week-old mice were obtained from Charles River Laboratories (France), and females were used for all experiments.…”

“…Parasites were harvested from the interface and collected by centrifugation at 500 g for 3 minutes. Gametocyte production and purification was performed as previously described [ 47 ]. Parasites were grown in mice that had been phenyl hydrazine-treated 3 days before infection.…”

Ca 2+ signals critical for egress and gametogenesis in malaria parasites depend on a multipass membrane protein that interacts with PKG

“…To further characterize the specificity of PbPKG and PbICM1 co-IPs in gametocytes, we compared the relative abundance of proteins recovered in both immunoprecipitates with those of seven other proteins involved in gametogenesis (21). Nine-dimensional principal components analysis (PCA) of all detected proteins confirmed the clustering of PbPKG and PbICM1, consistent with the formation of a complex (Fig.…”

“…Samples for excision PCRs were collected 24 hours after RAP treatment. P. berghei maintenance, purification, and transfection P. berghei maintenance, purification, and transfection were performed as previously described (21). P. berghei ANKA strain (36) derived clones 2.34 (27) and 615 ( 35), together with derived transgenic lines, were grown and maintained in CD1 outbred mice.…”