Pf CDPK1 is critical for malaria parasite gametogenesis and mosquito infection

Abstract

Efforts to knock out Plasmodium falciparum calcium-dependent protein kinase 1 (PfCDPK1) from asexual erythrocytic stage have not been successful, indicating an indispensable role of the enzyme in asexual growth. We recently reported generation of a transgenic parasite with mutant CDPK1 [Bansal A, et al. (2016) MBio 7:e02011-16]. The mutant CDPK1 (T145M) had reduced activity of transphosphorylation. We reasoned that CDPK1 could be disrupted in the mutant parasites. Consistent with this assumption, CDPK1 was successfully disrupted in the mutant parasites using CRISPR/Cas9. We and others could not disrupt PfCDPK1 in the WT parasites. The CDPK1 KO parasites show a slow growth rate compared with the WT and the CDPK1 T145M parasites. Additionally, the CDPK1 KO parasites show a defect in both male and female gametogenesis and could not establish an infection in mosquitoes. Complementation of the KO parasite with full-length PfCDPK1 partially rescued the asexual growth defect and mosquito infection. Comparative global transcriptomics of WT and the CDPK1 KO schizonts using RNA-seq show significantly high transcript expression of gametocyte-specific genes in the CDPK1 KO parasites. This study conclusively demonstrates that CDPK1 is a good target for developing transmission-blocking drugs.PfCDPK1 | plasmodium | compensation | gametocytes | mosquito

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“…The screen identified other known kinase inhibitors. MMV030734 is a specific inhibitor of Plasmodium calcium-dependent protein kinase 1 (CDPK1) (30), which is critical for parasite gametogenesis (31). MMV676182 and MMV688853 are bumped kinase inhibitors active against Toxoplasma and Cryptosporidium CDPK1 (32).…”

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“…The plasmodial orthologue of RAF kinase inhibitory protein (RKIP), referred to as PfPE‐PB1, has been shown to regulate the activity of P. falciparum calcium‐dependent protein kinase 1 ( Pf CDPK1) . Furthermore, disruption of the function of inhibition of Pf CDPK1 results in a multistage breakdown of the malarial life cycle . Sorafenib ( 561 ), which inhibits both the mutated and wild‐type human B‐Raf, has also been shown to inhibit P. falciparum viability and bind to Pf CDPK1, suggesting that inhibitors of human RAF orthologues may have utility as inhibitors of Pf CDPK1.…”

Plasmodial Kinase Inhibitors: License to Cure?

“… 40 Recently, Bansal et al were unable to create a knockout (KO) with a wild-type strain but were able to do so with a transgenic line containing a CDPK1 mutant (T145M). 38 The phenotype showed slow asexual proliferation compared to the wildtype indicating some degree of ambiguity around the essentiality of Pf CDPK1. Interestingly, the CDPK1 KO parasites are defective in the formation of male and female gametes and failed to establish infection in mosquitoes making Pf CDPK1 an interesting target for transmission blocking.…”