33 research outputs found
Kinesin-8B controls basal body function and flagellum formation and is key to malaria transmission
Eukaryotic flagella are conserved microtubule-based organelles
that drive cell motility. Plasmodium, the causative agent of
malaria, has a single flagellate stage: the male gamete in the
mosquito. Three rounds of endomitotic division in male gametocyte
together with an unusual mode of flagellum assembly
rapidly produce eight motile gametes. These processes are tightly
coordinated, but their regulation is poorly understood. To understand
this important developmental stage, we studied the
function and location of the microtubule-based motor kinesin-
8B, using gene-targeting, electron microscopy, and live cell
imaging. Deletion of the kinesin-8B gene showed no effect on
mitosis but disrupted 9+2 axoneme assembly and flagellum
formation during male gamete development and also completely
ablated parasite transmission. Live cell imaging showed that
kinesin-8B–GFP did not co-localise with kinetochores in the
nucleus but instead revealed a dynamic, cytoplasmic localisation
with the basal bodies and the assembling axoneme during flagellum
formation. We, thus, uncovered an unexpected role for
kinesin-8B in parasite flagellum formation that is vital for the
parasite life cycle
Plasmodium kinesin-8X associates with mitotic spindles and is essential for oocyst development during parasite proliferation and transmission
Kinesin-8 proteins are microtubule motors that are often involved in regulation of mitotic spindle length and chromosome alignment. They move towards the plus ends of spindle microtubules and regulate the dynamics of these ends due, at least in some species, to their microtubule depolymerization activity. Plasmodium spp. exhibit an atypical endomitotic cell division in which chromosome condensation and spindle dynamics in the different proliferative stages are not well understood. Genome-wide shared orthology analysis of Plasmodium spp. revealed the presence of two kinesin-8 motor proteins, kinesin-8X and kinesin-8B. Here we studied the biochemical properties of kinesin-8X and its role in parasite proliferation. In vitro, kinesin-8X has motility and depolymerization activities like other kinesin-8 motors. To understand the role of Plasmodium kinesin-8X in cell division, we used fluorescence-tagging and live cell imaging to define its location, and gene targeting to analyse its function, during all proliferative stages of the rodent malaria parasite P. berghei life cycle. The results revealed a spatio-temporal involvement of kinesin-8X in spindle dynamics and an association with both mitotic and meiotic spindles and the putative microtubule organising centre (MTOC). Deletion of the kinesin-8X gene revealed a defect in oocyst development, confirmed by ultrastructural studies, suggesting that this protein is required for oocyst development and sporogony. Transcriptome analysis of Δkinesin-8X gametocytes revealed modulated expression of genes involved mainly in microtubule-based processes, chromosome organisation and the regulation of gene expression, supporting a role for kinesin-8X in cell division. Kinesin-8X is thus required for parasite proliferation within the mosquito and for transmission to the vertebrate host