Abstract
During cell division microtubules capture chromosomes by binding to the kinetochore assembled in the centromeric region of chromosomes. In mitosis sister chromatids are captured by microtubules emanating from both spindle poles, a process called bipolar attachment, whereas in meiosis I sisters are attached to microtubules originating from one spindle pole, called monopolar attachment. For determining chromosome orientation, kinetochore geometry or structure might be an important target of regulation. However, the molecular basis of this regulation has remained elusive. Here we show the link between kinetochore orientation and cohesion within the centromere in fission yeast Schizosaccharomyces pombe by strategies developed to visualize the concealed cohesion within the centromere, and to introduce artificial tethers that can influence kinetochore geometry. Our data imply that cohesion at the core centromere induces the mono-orientation of kinetochores whereas cohesion at the peri-centromeric region promotes bi-orientation. Our study may reveal a general mechanism for the geometric regulation of kinetochores, which collaborates with previously defined tension-dependent reorientation machinery.
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Acknowledgements
We thank S. Hauf for critically reading the manuscript. We thank H. Matsuzaki, A. Yamamoto, R. Allshire, K. Nasmyth and the Yeast Genetic Resource Center (YGRC) for yeast strains. We also thank all the members of our laboratory for their support and discussion, especially S. Yokobayashi for materials and assistance in the initial stage of this project. This work was supported in part by Special Coordination Funds for Promoting Science and Technology (to T.S.), the Global COE Program (Integrative Life Science Based on the Study of Biosignaling Mechanisms), MEXT, Japan, and a Grant-in-Aid for Specially Promoted Research from the Ministry of Education, Culture, Sports, Science and Technology of Japan (to Y.W.).
Author Contributions T.S. and Y.W. conceived and designed the experiments. T.S. performed all experiments. K.T. set up the TetR–tdTomato system in fission yeast. Y.W. planned research and wrote the manuscript with input from T.S.
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Supplementary Information
This file contains Supplementary Table 1, Supplementary Figures 1-13 with Legends and Supplementary References. (PDF 3130 kb)
Supplementary Movie 1
This movie shows wild-type prophase I zygote classified as "No exc." in Fig.1b. (MOV 122 kb)
Supplementary Movie 2
This movie shows wild-type prophase I zygote classified as "One exc." in Fig.1b. (MOV 126 kb)
Supplementary Movie 3
This movie shows wild-type prophase I zygote classified as "Cohered" in Fig.1b. (MOV 91 kb)
Supplementary Movie 4
This movie shows wild-type prophase I zygote classified as "Separated" in Fig.1b. (MOV 112 kb)
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Sakuno, T., Tada, K. & Watanabe, Y. Kinetochore geometry defined by cohesion within the centromere. Nature 458, 852–858 (2009). https://doi.org/10.1038/nature07876
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DOI: https://doi.org/10.1038/nature07876
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