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Molecular mechanisms of kinetochore capture by spindle microtubules


For high-fidelity chromosome segregation, kinetochores must be properly captured by spindle microtubules, but the mechanisms underlying initial kinetochore capture have remained elusive. Here we visualized individual kinetochore–microtubule interactions in Saccharomyces cerevisiae by regulating the activity of a centromere. Kinetochores are captured by the side of microtubules extending from spindle poles, and are subsequently transported poleward along them. The microtubule extension from spindle poles requires microtubule plus-end-tracking proteins and the Ran GDP/GTP exchange factor. Distinct kinetochore components are used for kinetochore capture by microtubules and for ensuring subsequent sister kinetochore bi-orientation on the spindle. Kar3, a kinesin-14 family member, is one of the regulators that promote transport of captured kinetochores along microtubules. During such transport, kinetochores ensure that they do not slide off their associated microtubules by facilitating the conversion of microtubule dynamics from shrinkage to growth at the plus ends. This conversion is promoted by the transport of Stu2 from the captured kinetochores to the plus ends of microtubules.

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Figure 1: Visualizing kinetochore capture and transport along microtubules.
Figure 2: Mechanisms for nuclear microtubule extension and for kinetochore capture.
Figure 3: Captured kinetochores facilitate microtubule rescue.
Figure 4: Microtubule rescue at the plus end coincides with the arrival of Stu2 protein transported from captured kinetochores.
Figure 5: Kar3 kinesin is involved in the poleward transport of kinetochores along the side of microtubules.
Figure 6: Association of authentic centromeres with microtubules in unperturbed cell cycles.


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We thank J. R. Swedlow, M. J. R. Stark, A. Gartner, M. A. Hoyt, A. Desai, P. R. Clarke, P. D. Andrews and members of the Tanaka laboratory for discussions and for reading the manuscript; T. Hyman, K. Nasmyth, I. W. Mattaj, E. Karsenti, F. Uhlmann and J. Ellenberg for discussions; J.-F. Maure, N. Rachidi and M. J. R. Stark for sharing their unpublished data; Y. Kitamura, S. Swift, M. Romao and G. Keir for technical help; F. Wheatley and the media kitchen for media preparation; M. A. Hoyt, D. Pellman, R. Ciosk, F. Uhlmann, K. Nasmyth, T. C. Huffaker, J. V. Kilmartin, E. Schiebel, S. Biggins, C. S. M. Chan, I. M. Cheeseman, G. Barnes, R. Tsien, S. J. Elledge, J. Lechner, A. H. Corbett, P. A. Silver, P. K. Sorger, X. He, A. F. Straight, M. D. Rose, V. Doye, F. Severin, I. Ouspenski, K. Bloom, T. Nishimoto, J. E. Haber, T. N. Davis, EUROSCARF and the Yeast Resource Center for reagents. This work was supported by The Wellcome Trust, Cancer Research UK and the EMBO Young Investigator Program.

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Supplementary information

Supplementary Notes

This file contains Supplementary Notes and Supplementary Figures S1-S13. (PDF 462 kb)

Supplementary Video Legends

Legends to accompany the below Supplementary Videos. (RTF 4 kb)

Supplementary Video S1

Video of the cell shown in Fig 1b. Speed in the video is 100 times faster than the actual motion. (MOV 6892 kb)

Supplementary Video S2

Video of the cell shown in Fig 1c. Speed in the video is 100 times faster than the actual motion. (MOV 6034 kb)

Supplementary Video S3

Video of the cell shown in Supplementary Fig S2. Speed in the video is 100 times faster than the actual motion. (MOV 3798 kb)

Supplementary Video S4

Video of the cell shown in Supplementary Fig S4a. Speed in the video is 100 times faster than the actual motion. (MOV 1839 kb)

Supplementary Video S5

Video of the cell shown in Fig 3a, top. Speed in the video is 100 times faster than the actual motion. (MOV 2663 kb)

Supplementary Video S6

Video of the cell shown in Fig 3a, bottom. Speed in the video is 100 times faster than the actual motion. (MOV 5923 kb)

Supplementary Video S7

Video of the cells shown in Fig 4a. Speed in the video is 50 times faster than the actual motion. (MOV 5193 kb)

Supplementary Video S8

Video of a wild type control for Supplementary Video S9 and S10. This cell is not shown in any panels of figures. KAR3+ cells (T3531) were treated as in Fig 5. GFP and YFP signals were collected together as in Supplementary Video S9 and S10). Speed in the video is 300 times faster than the actual motion. (MOV 2532 kb)

Supplementary Video S9

Video of the kar3-1 cell shown in Fig 5a. Speed in the video is 300 times faster than the actual motion. (MOV 2601 kb)

Supplementary Video S10

Video of the KAR3-overexpressed cell shown in Fig 5a. Speed in the video is 300 times faster than the actual motion. (MOV 2460 kb)

Supplementary Video S11

Video of the cell shown in Supplementary Fig S12. Speed in the video is 150 times faster than the actual motion. (MOV 1603 kb)

Video S12

Video of the cell shown in Fig 6a. Speed in the video is 50 times faster than the actual motion. (MOV 4278 kb)

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Tanaka, K., Mukae, N., Dewar, H. et al. Molecular mechanisms of kinetochore capture by spindle microtubules. Nature 434, 987–994 (2005).

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