Mechanics of microtubule organizing center clustering and spindle positioning in budding yeast Cryptococcus neoformans

The dynamic process of mitotic spindle assembly depends on multitudes of inter-dependent interactions involving kinetochores (KTs), microtubules (MTs), spindle pole bodies (SPBs), and molecular motors. Before forming the mitotic spindle, multiple visible microtubule organizing centers (MTOCs) coalesce into a single focus to serve as an SPB in the pathogenic budding yeast, Cryptococcus neoformans. To explain this unusual phenomenon in the fungal kingdom, we propose a search and capture model, in which cytoplasmic MTs (cMTs) nucleated by MTOCs grow and capture each other to promote MTOC clustering. Our quantitative modeling identifies multiple redundant mechanisms mediated by a combination of cMT-cell cortex interactions and inter-cMT coupling to facilitate MTOC clustering within the physiological time limit as determined by timelapse live-cell microscopy. Besides, we screen various possible mechanisms by computational modeling and propose optimal conditions that favor proper spindle positioning-a critical determinant for timely chromosome segregation. These analyses also reveal that a combined effect of MT buckling, dynein pull, and cortical push maintains spatiotemporal spindle localization.

Automated summary

The dynamic process of mitotic spindle assembly is complex and involves multiple inter-dependent interactions. In the fungal kingdom, multiple visible microtubule organizing centers coalesce into a single focus before forming the mitotic spindle, promoting MTOC clustering through a search and capture model. Various mechanisms, including interactions between cMTs and the cell cortex, as well as inter-cMT coupling, facilitate MTOC clustering within a physiological time limit.