Real-time dynamics of Plasmodium NDC80 as a marker for the kinetochore during atypical mitosis and meiosis
Pandey R., Zeeshan M., Ferguson DJP., Markus R., Brady D., Daniel E., Stanway R., Holder A., Guttery D., Tewari R.
Abstract Eukaryotic cell proliferation requires chromosome replication and precise segregation, followed by cell division, to ensure daughter cells have identical genomic copies. The kinetochore is a multi-protein structure assembled on chromosome centromeres, which mediates attachment to spindle microtubules and chromosome segregation during mitosis. Plasmodium spp ., the causative agent of malaria, undergoes closed mitosis with an intact nuclear membrane and intranuclear mitotic spindle. However, the regulation of the main events of mitosis and meiosis, including chromosome segregation, is poorly understood in this parasite. In part this is due to a distinct lack of markers for tracking kinetochore dynamics. Since NDC80 is the conserved component of the kinetochore complex, we labelled NDC80 with either a C-terminal GFP or mCherry tag by recombination at the endogenous locus to generate transgenic lines of the rodent malaria parasite Plasmodium berghei . We show that NDC80 is located at a single cluster of all kinetochores and examine its spatio-temporal dynamics throughout all proliferative stages of the parasite life cycle. We also compare the location of NDC80 with that of the kinetochore using electron microscopy. These transgenic parasite lines are a very useful resource to identify and study the kinetochore complex and follow chromosome dynamics during malaria parasite development. Summary Statement The dynamic localization of NDC80, a kinetochore marker, is examined throughout the proliferative stages of malaria parasite life-cycle which provides a useful resource to study kinetochore complexes and chromosome dynamics.