Correlation between Cyclin Dependent Kinases and Artemisinin-Induced Dormancy in Plasmodium falciparum In Vitro

Gray, Karen-Ann; Gresty, Karryn; Chen, Nanhua; Zhang, Veronica; Gutteridge, Clare E.; Peatey, Christopher L.; Chavchich, Marina; Waters, Norman C.; Cheng, Qin

doi:10.1371/journal.pone.0157906

Cited by 19 publications

(19 citation statements)

References 50 publications

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“…The peculiarities of Plasmodium schizogony begin with the lack of a G2 phase as the syncytial nuclei appear to alternate asynchronously between S and M phases prior to the orchestrated event of cytokinesis [ 35 ] (Fig. 4a ).…”

Section: Regulation Of the Plasmodium Cell Cyclementioning

confidence: 99%

Checks and balances? DNA replication and the cell cycle in Plasmodium

2018

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It is over 100 years since the life-cycle of the malaria parasite Plasmodium was discovered, yet its intricacies remain incompletely understood - a knowledge gap that may prove crucial for our efforts to control the disease. Phenotypic screens have partially filled the void in the antimalarial drug market, but as compound libraries eventually become exhausted, new medicines will only come from directed drug development based on a better understanding of fundamental parasite biology. This review focusses on the unusual cell cycles of Plasmodium, which may present a rich source of novel drug targets as well as a topic of fundamental biological interest. Plasmodium does not grow by conventional binary fission, but rather by several syncytial modes of replication including schizogony and sporogony. Here, we collate what is known about the various cell cycle events and their regulators throughout the Plasmodium life-cycle, highlighting the differences between Plasmodium, model organisms and other apicomplexan parasites and identifying areas where further study is required. The possibility of DNA replication and the cell cycle as a drug target is also explored. Finally the use of existing tools, emerging technologies, their limitations and future directions to elucidate the peculiarities of the Plasmodium cell cycle are discussed.

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Section: Regulation Of the Plasmodium Cell Cyclementioning

confidence: 99%

Checks and balances? DNA replication and the cell cycle in Plasmodium

2018

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“…The increased enrichment preference for live, quiescent parasites over dead parasites is likely due to the loss of cholesterol metabolism in parasites upon death. While quiescent parasites down-regulate many aspects of metabolism 31 , these data indicate that quiescent parasites continue the utilization of cholesterol.…”

Section: Discussionmentioning

confidence: 87%

“…7a, SYBR-Green/MitoTracker Red positive). When compared to untreated rings, quiescent parasites showed reduced MMP staining area, indicative of the condensed cytoplasm and reduced metabolism seen in quiescence 31,32 . While quiescent parasites were enriched by the removal of uninfected erythrocyte material, erythrocytes containing dead parasites were also enriched by this process (Fig.…”

Section: Slope Enriches In Vitro-derived Quiescent Parasitesmentioning

confidence: 96%

Cholesterol-dependent enrichment of understudied erythrocytic stages of human Plasmodium parasites

Brown

Moore

Guler

2020

Sci Rep

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for intracellular pathogens, the host cell provides needed protection and nutrients. A major challenge of intracellular parasite research is collection of high parasite numbers separated from host contamination. This situation is exemplified by the malaria parasite, which spends a substantial part of its life cycle inside erythrocytes as rings, trophozoites, and schizonts, before egress and reinvasion. erythrocytic Plasmodium parasite forms refractory to enrichment remain understudied due to high host contamination relative to low parasite numbers. Here, we present a method for separating all stages of Plasmodium-infected erythrocytes through lysis and removal of uninfected erythrocytes. The Streptolysin O-Percoll (SLOPE) method is effective on previously inaccessible forms, including circulating rings from malaria-infected patients and artemisinin-induced quiescent parasites. SLope can be used on multiple parasite species, under multiple media formulations, and lacks measurable impacts on parasite viability. We demonstrate erythrocyte membrane cholesterol levels modulate the preferential lysis of uninfected host cells by SLO, and therefore modulate the effectiveness of SLOPE. Targeted metabolomics of SLOPE-enriched ring stage samples confirms parasite-derived metabolites are increased and contaminating host material is reduced compared to non-enriched samples. Due to consumption of cholesterol by other intracellular bacteria and protozoa, SLope holds potential for improving research on organisms beyond Plasmodium. Malaria, caused by protozoan parasites of the Plasmodium genus, is a continuing threat to global health. A total of five Plasmodium species cause malaria in humans, with Plasmodium falciparum being responsible for the large majority of malaria morbidity and mortality 1. While the global malaria burden has decreased over the past decade, the emergence and spread of antimalarial resistant Plasmodium threatens to undo this progress and emphasizes the dire need to understand more about the biology of this parasite. The current World Health Organization recommendation for treatment of malaria is artemisinin combination therapy 2. However, clinical resistance has now been reported to both artemisinin and almost all of its partner drugs 3-5. All symptoms of malaria, including cyclical fevers and hypoglycemia, occur due to the asexual replication cycle of the parasite within human erythrocytes (Fig. 1a). Parasites undergo rounds of replication progressing from the ring stage, to trophozoites and schizonts, before rupturing from host erythrocytes to release merozoites, which go on to invade new erythrocytes and continue the cycle of infection 6. Many studies aiming to understand the biology of asexual Plasmodium are performed only on late stage parasite samples (trophozoites and schizonts). This is due in part to the larger biomass of these stages but also to the existence of effective enrichment methods 7 ; erythrocytes infected with late stage parasites can be separated from uninfected erythrocytes by density...

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“…In the schizont stage, the inhibition of P. falciparum Cdc2-related kinase-1(PfCRK-1) protein interferes with the growth of P. falciparum [30], and the depletion of Plasmodium-speci c kinase PfCRK4 result in the complete blockage of nuclear division and intensively inhibits DNA replication [31]. The downregulation of P. falciparum MO15-related protein kinase (PfMRK), Cdc2-related kinase PfPK5, P. falciparum CRK3 and other cyclins during dormancy arrests parasite progression at the G1 phase, and halts DNA synthesis [32]. The inhibition of CDK7 kinase causes transcript dysregulation in T. gondii [33].…”

Section: Discussionmentioning

confidence: 99%

Diclazuril-Induced Expression of CDK-Related Kinase 2 in the Second-Generation Merozoites of Eimeria Tenella

Zhou

Ding

Yang

et al. 2020

Preprint

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Background：Coccidiosis caused by Eimeria tenella infection, directly or indirectly leads to great loss to poultry industry. With the emergence of drug-resistance in chicken coccidia, it is imperative to develop new drugs. Cyclin-dependent kinases (CDKs) regulate cell cycle progression in numerous organisms by acting as key molecular switches. Results: In the present study, a diclazuril anticoccidiosis animal model was established and CDK-related kinase 2 (CRK2) in the second-generation merozoite of E. tenella (EtCRK2) gene was amplified through reverse transcription-polymerase chain reaction (RT-PCR) and expressed in Escherichia coli Rosetta (DE3). Purified recombinant protein was used for antiserum preparation. Subsequently, EtCRK2 transcription and translation levels were detected through quantitative real-time PCR and Western blot analysis, respectively. The localization of EtCRK2 in merozoites was examined via immunofluorescence techniques. Results showed that the mRNA and protein expression levels of EtCRK2 decreased in the infected/diclazuril group compared with those in the infected/control group. In addition, immunofluorescence analysis showed that EtCRK2 was localized in the cytoplasm of merozoites. The fluorescence intensity of EtCRK2 in the infected/diclazuril group was significantly weaker than that in the infected/control group. Conclusions: This study demonstrated that the anticoccidial drug diclazuril against E. tenella by affecting the expression pattern of EtCRK2 molecule, and EtCRK2 may be used as a candidate target for new drug development.

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Correlation between Cyclin Dependent Kinases and Artemisinin-Induced Dormancy in Plasmodium falciparum In Vitro

Cited by 19 publications

References 50 publications

Checks and balances? DNA replication and the cell cycle in Plasmodium

Checks and balances? DNA replication and the cell cycle in Plasmodium

Cholesterol-dependent enrichment of understudied erythrocytic stages of human Plasmodium parasites

Diclazuril-Induced Expression of CDK-Related Kinase 2 in the Second-Generation Merozoites of Eimeria Tenella

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