Abstract
Proper chromosome segregation is crucial for cell division. In eukaryotes, this is achieved by the kinetochore, an evolutionarily conserved multi-protein complex that physically links the DNA to spindle microtubules and takes an active role in monitoring and correcting erroneous spindle-chromosome attachments. Our mechanistic understanding of these functions and how they ensure an error-free outcome of mitosis is still limited, partly because we lack a comprehensive understanding of the kinetochore structure in the cell. In this study, we use single-molecule localization microscopy to visualize individual kinetochore complexes in situ in budding yeast. For major kinetochore proteins, we measured their abundance and position within the metaphase kinetochore. Based on this comprehensive dataset, we propose a quantitative model of the budding yeast kinetochore. While confirming many aspects of previous reports based on bulk imaging, our results present a unifying nanoscale model of the kinetochore in budding yeast.
Competing Interest Statement
The authors have declared no competing interest.
Footnotes
To address the reviewers' points, we now quantified the measurement errors (new Fig. S1 and Fig. S6) and refined the localization filtering accordingly. All the affected results were now updated (Fig. 2, Fig. 3, Fig. 4, Table 1, Fig. S7, and Table S3) and showed no significant difference compared to the last version. We also included new results of Ask1, a subunit of the Dam1 complex. Our structural model of the budding yeast kinetochore was now updated (Fig. 4). A correlation-based analysis was now included (new Fig. S5). Additional information from relevant works and this work was now provided (new Fig. S4, Table S1, and Table S4). Texts were updated to reflect these changes and for clarity.