Real-time dynamics of <i>Plasmodium</i> NDC80 reveals unusual modes of chromosome segregation during parasite proliferation

Pandey, Rajan Kumar; Zeeshan, Mohammad; Ferguson, David J. P.; Márkus, Róbert; Brady, Declan; Daniel, Emilie; Stanway, Rebecca R.; Holder, Anthony A.; Guttery, David S.; Tewari, Rita

doi:10.1101/767830

Cited by 11 publications

(32 citation statements)

References 65 publications

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“…5B). This mirrors what has been seen in electron microscopy studies: nuclear spindles radiate from the nuclear poles, with attached kinetochores within an intact nuclear membrane during oocyst development [30,32].…”

Section: Pbkinesin-5-gfp Exhibits Multiple Nuclear Foci During Oocystsupporting

confidence: 78%

“…4D). This result showed that kinesin-5 distribution and localization is associated with spindle dynamics, similar to the behaviour of NDC80 [32], and can be blocked by taxol treatment, which binds tubulin and stabilises MTs by preventing depolymerisation (Fig. 4D).…”

Section: Spatiotemporal Dynamics Of Pbkinesin-5 Reveal Its Location Osupporting

confidence: 63%

“…Each oocyst contains multiple lobes and produces thousands of sporozoites [32]. Kinesin-5-GFP fluorescence was observed as multiple foci representing a location at the putative MTOC/nuclear poles, from very early in development (day 7) to late stages (at day 14) of oocyst maturation (Fig.…”

Section: Pbkinesin-5-gfp Exhibits Multiple Nuclear Foci During Oocystmentioning

confidence: 97%

“…The Plasmodium life cycle has two unusual mitotic processes that occur during schizogony/sporogony and male gametogony, and a single meiotic stage during zygote to ookinete transformation [32]. To examine any functional role of kinesin-5 during these processes, we deleted the gene from the P. berghei genome using a double crossover homologous recombination strategy in a parasite line constitutively expressing green fluorescent protein (GFP) at all stages of the parasite life cycle ( Fig.…”

Section: Pbkinesin-5 Is Dispensable For Cell Division During the Diffmentioning

confidence: 99%

“…To further investigate whether the location of kinesin-5 is cytoplasmic or nuclear, the kinesin-5-GFP parasite line was genetically crossed with the NDC80-mCherry kinetochore marker line [32] and kinesin-8B-mCherry cytoplasmic axoneme marker line [23], and the crosses were used for live cell imaging of both markers to establish their spatiotemporal relationship.…”

Section: Spatiotemporal Dynamics Of Pbkinesin-5 Reveal Its Location Omentioning

confidence: 99%

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Plasmodium bergheikinesin-5 associates with the spindle apparatus during cell division and is important for efficient production of infectious sporozoites

Zeeshan

Brady

Stanway

et al. 2020

Preprint

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AbstractKinesin-5 motors play essential roles in spindle apparatus assembly during cell division, by generating forces to establish and maintain the spindle bipolarity essential for proper chromosome segregation. Kinesin-5 is largely conserved structurally and functionally in model eukaryotes, but its role is unknown in the Plasmodium parasite, an evolutionarily divergent organism with several atypical features of both mitotic and meiotic cell division. We have investigated the function and subcellular location of kinesin-5 during cell division throughout the Plasmodium berghei life cycle. Deletion of kinesin-5 had little visible effect at any proliferative stage. The only significant effect was a decrease in the number of motile sporozoites in mosquito salivary glands, which were able to infect a new vertebrate host. Live-cell imaging showed kinesin-5-GFP located on the spindle and at spindle poles during both atypical mitosis and meiosis. Fixed-cell immunofluorescence assays revealed kinesin-5 co-localized with α-tubulin and centrin-2 and exhibited a partial overlap with kinetochore marker NDC80 during early blood stage schizogony. Dual-colour live-cell imaging during male gametogony showed that kinesin-5 is closely associated with NDC80, but not with kinesin-8B, a marker of the basal body and axonemes of the forming flagella. Treatment of gametocytes with microtubule-specific inhibitors confirmed kinesin-5 association with nuclear spindles and not cytoplasmic axonemal microtubules. Altogether, our results demonstrate that kinesin-5 is associated with the spindle apparatus and expressed in proliferating parasite stages, but it is not essential for parasite survival.ImportanceKinesin-5 is a motor protein with important roles during cell proliferation and therefore considered a strong target for therapeutics development against many diseases. The role of kinesin-5 in Plasmodium is unknown, but in this organism there are atypical aspects of cell division that differ from those of model eukaryotes. For example, classical regulators such as polo-like kinase, classical cyclins and cyclin dependent kinases are absent from Plasmodium. Here, we show that in Plasmodium, unlike in most model eukaryotes, kinesin-5 is dispensable, suggesting that the parasite uses somewhat different machinery for cell proliferation. The study is important because it shows that kinesin-5 is not essential universally for cell division and suggests that there may be other non-canonical mechanisms and regulators. The general concept that kinesin-5 is essential for spindle pole formation and chromosome segregation during cell division is not true for Plasmodium, and this is of interest to a broad readership.

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Section: Pbkinesin-5-gfp Exhibits Multiple Nuclear Foci During Oocystsupporting

confidence: 78%

Section: Spatiotemporal Dynamics Of Pbkinesin-5 Reveal Its Location Osupporting

confidence: 63%

Section: Pbkinesin-5-gfp Exhibits Multiple Nuclear Foci During Oocystmentioning

confidence: 97%

Section: Pbkinesin-5 Is Dispensable For Cell Division During the Diffmentioning

confidence: 99%

Section: Spatiotemporal Dynamics Of Pbkinesin-5 Reveal Its Location Omentioning

confidence: 99%

See 3 more Smart Citations

Plasmodium bergheikinesin-5 associates with the spindle apparatus during cell division and is important for efficient production of infectious sporozoites

Zeeshan

Brady

Stanway

et al. 2020

Preprint

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Repurposing the mitotic machinery to drive cellular elongation and chromatin reorganisation in Plasmodium falciparum gametocytes

Shami

Cho

et al. 2022

Nat Commun

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The sexual stage gametocytes of the malaria parasite, Plasmodium falciparum, adopt a falciform (crescent) shape driven by the assembly of a network of microtubules anchored to a cisternal inner membrane complex (IMC). Using 3D electron microscopy, we show that a non-mitotic microtubule organizing center (MTOC), embedded in the parasite’s nuclear membrane, orients the endoplasmic reticulum and the nascent IMC and seeds cytoplasmic microtubules. A bundle of microtubules extends into the nuclear lumen, elongating the nuclear envelope and capturing the chromatin. Classical mitotic machinery components, including centriolar plaque proteins, Pfcentrin-1 and −4, microtubule-associated protein, End-binding protein-1, kinetochore protein, PfNDC80 and centromere-associated protein, PfCENH3, are involved in the nuclear microtubule assembly/disassembly process. Depolymerisation of the microtubules using trifluralin prevents elongation and disrupts the chromatin, centromere and kinetochore organisation. We show that the unusual non-mitotic hemispindle plays a central role in chromatin organisation, IMC positioning and subpellicular microtubule formation in gametocytes.

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Repurposing of synaptonemal complex proteins for kinetochores in Kinetoplastida

et al. 2021

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Chromosome segregation in eukaryotes is driven by the kinetochore, a macromolecular complex that connects centromeric DNA to microtubules of the spindle apparatus. Kinetochores in well-studied model eukaryotes consist of a core set of proteins that are broadly conserved among distant eukaryotic phyla. By contrast, unicellular flagellates of the class Kinetoplastida have a unique set of 36 kinetochore components. The evolutionary origin and history of these kinetochores remain unknown. Here, we report evidence of homology between axial element components of the synaptonemal complex and three kinetoplastid kinetochore proteins KKT16-18. The synaptonemal complex is a zipper-like structure that assembles between homologous chromosomes during meiosis to promote recombination. By using sensitive homology detection protocols, we identify divergent orthologues of KKT16-18 in most eukaryotic supergroups, including experimentally established chromosomal axis components, such as Red1 and Rec10 in budding and fission yeast, ASY3-4 in plants and SYCP2-3 in vertebrates. Furthermore, we found 12 recurrent duplications within this ancient eukaryotic SYCP 2–3 gene family, providing opportunities for new functional complexes to arise, including KKT16-18 in the kinetoplastid parasite Trypanosoma brucei . We propose the kinetoplastid kinetochore system evolved by repurposing meiotic components of the chromosome synapsis and homologous recombination machinery that were already present in early eukaryotes.

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Real-time dynamics of Plasmodium NDC80 reveals unusual modes of chromosome segregation during parasite proliferation

Cited by 11 publications

References 65 publications

Plasmodium bergheikinesin-5 associates with the spindle apparatus during cell division and is important for efficient production of infectious sporozoites

Plasmodium bergheikinesin-5 associates with the spindle apparatus during cell division and is important for efficient production of infectious sporozoites

Repurposing the mitotic machinery to drive cellular elongation and chromatin reorganisation in Plasmodium falciparum gametocytes

Repurposing of synaptonemal complex proteins for kinetochores in Kinetoplastida

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