Plasmodium kinesin-8X associates with mitotic spindles and is essential for oocyst development during parasite proliferation and transmission

Zeeshan, Mohammad; Shilliday, Fiona; Liu, Tianyang; Abel, Steven; Mourier, Tobias; Ferguson, David J P; Rea, Edward; Stanway, Rebecca R; Roques, Magali; Williams, Desiree; Daniel, Emilie; Brady, Declan; Roberts, Anthony J; Holder, Anthony A; Pain, Arnab; Le, Karine G; Le Roch, Roch G; Moores, Carolyn A; Tewari, Rita

Abstract

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.

Citation

Zeeshan, M., Shilliday, F., Liu, T., Abel, S., Mourier, T., Ferguson, D. J. P., Rea, E., Stanway, R. R., Roques, M., Williams, D., Daniel, E., Brady, D., Roberts, A. J., Holder, A. A., Pain, A., Le, K. G., Le Roch, R. G., Moores, C. A., & Tewari, R. (2019). Plasmodium kinesin-8X associates with mitotic spindles and is essential for oocyst development during parasite proliferation and transmission. PLoS Pathogens, 15(10), Article e1008048. https://doi.org/10.1371/journal.ppat.1008048

• Journal Article Type: Article
• Acceptance Date: Aug 26, 2019
• Online Publication Date: Oct 10, 2019
• Publication Date: Oct 1, 2019
• Deposit Date: Sep 30, 2019
• Publicly Available Date: Oct 1, 2019
• Journal: PLoS pathogens
• Print ISSN: 1553-7366
• Electronic ISSN: 1553-7374
• Publisher: Public Library of Science
• Peer Reviewed: Peer Reviewed
• Volume: 15
• Issue: 10
• Article Number: e1008048
• DOI: https://doi.org/10.1371/journal.ppat.1008048
• Keywords: Immunology; Genetics; Molecular Biology; Microbiology; Parasitology; Virology
• Public URL: https://nottingham-repository.worktribe.com/output/2723065
• Contract Date: Oct 1, 2019