DNA replication dynamics during erythrocytic schizogony in the malaria parasites Plasmodium falciparum and Plasmodium knowlesi

Author summaryMalaria parasites are unusual, early-diverging single-celled organisms. One of their atypical features is their mode of producing new cells. Most cells replicate their genome, segregate the copies into two nuclei and then split the whole cell in two: a process called binary fission. Malaria parasites, by contrast, multiply primarily by schizogony. Each cell replicates its genome several times over, asynchronously, generating many nuclei within the same cytoplasm, before splitting into many new daughter cells in a single mass event. All malaria species do this, but there are stark differences between species in how long schizogony takes and how many progeny each cell can produce. Understanding this is important because the rate of parasite reproduction is fundamentally important to malarial disease: humans who have a high burden of parasites in their blood are most likely to suffer severe malaria. Here we compare the process of schizogony in two different species, by developing cell lines in which we can follow de novo DNA replication at high spatial and temporal resolution, at both whole-cell and single-molecule levels. We establish the dynamics of schizogony, highlighting similarities and differences between two species and setting parameters for future studies of interventions that could interfere with parasite reproduction. Malaria parasites are unusual, early-diverging protozoans with non-canonical cell cycles. They do not undergo binary fission, but divide primarily by schizogony. This involves the asynchronous production of multiple nuclei within the same cytoplasm, culminating in a single mass cytokinesis event. The rate and efficiency of parasite reproduction is fundamentally important to malarial disease, which tends to be severe in hosts with high parasite loads. Here, we have studied for the first time the dynamics of schizogony in two human malaria parasite species, Plasmodium falciparum and Plasmodium knowlesi. These differ in their cell-cycle length, the number of progeny produced and the genome composition, among other factors. Comparing them could therefore yield new information about the parameters and limitations of schizogony. We report that the dynamics of schizogony differ significantly between these two species, most strikingly in the gap phases between successive nuclear multiplications, which are longer in P. falciparum and shorter, but more heterogenous, in P. knowlesi. In both species, gaps become longer as schizogony progresses, whereas each period of active DNA replication grows shorter. In both species there is also extreme variability between individual cells, with some schizonts producing many more nuclei than others, and some individual nuclei arresting their DNA replication for many hours while adjacent nuclei continue to replicate. The efficiency of schizogony is probably influenced by a complex set of factors in both the parasite and its host cell.

Automated summary

This study compares the process of schizogony in two different human malaria parasite species and highlights the significant differences in nuclear replication gaps and nuclear numbers within cells. These differences are important for understanding the pathogenesis of malaria and for developing interventions to interfere with parasite reproduction.