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Cohesin ensures bipolar attachment of microtubules to sister centromeres and resists their precocious separation

Nature Cell Biology volume 2, pages 492–499 (2000)Cite this article

Abstract

The multisubunit protein complex cohesin is required to establish cohesion between sister chromatids during S phase and to maintain it during G2 and M phases. Cohesin is essential for mitosis, and even partial defects cause very high rates of chromosome loss. In budding yeast, cohesin associates with specific sites which are distributed along the entire length of a chromosome but are more dense in the vicinity of the centromere. Real-time imaging of individual centromeres tagged with green fluorescent protein suggests that cohesin bound to centromeres is important for bipolar attachment to microtubules. This cohesin is, however, incapable of resisting the consequent force, which leads to sister centromere splitting and chromosome stretching. Meanwhile, cohesin bound to sequences flanking the centromeres prevents sister chromatids from completely unzipping and is required to pull back together sister centromeres that have already split. Cohesin therefore has a central role in generating a dynamic tension between microtubules and sister chromatid cohesion at centromeres, which lasts until chromosome segregation is finally promoted by separin-dependent cleavage of the cohesin subunit Scc1p.

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Figure 1: Sister centromeres separate earlier than regions along chromosome arms, in a Cdc20p-independent manner.
Figure 2: Early separation of centromere regions in short-spindle cells detected by FISH.
Figure 3: Sister centromeres split and reassociate repeatedly before the onset of anaphase B.
Figure 4: Sister centromeres split and do not reassociate in the absence of Scc1p.
Figure 5: Both sister centromeres are frequently found moving in the vicinity of a single SPB in the absence of Scc1p.
Figure 6: Model illustrating cohesin distribution along chromosomes, bipolar attachment of kinetochores to microtubules, centromere movements and onset of anaphase.

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Acknowledgements

We especially thank K. Paiha, P. Steinlein, A. Desai and T. Hyman for helping to set up the time-lapse system. We also thank Q.-w. Jin, F. Severin, T. Hyman, M. Glotzer and people in the Nasmyth laboratory for encouraging discussions; R. Ciosk, F. Uhlmann and E. Schiebel for yeast strains and plasmids; G. Goshima, M. Yanagida, X. He and P. Sorger for communicating unpublished results; M. Glotzer, F. Uhlmann and V. Katis for their critical comments on the manuscript; and M. Galova, H. Tkadletz and J. Erdei for their technical help. This work was supported by grants from the Austrian Industrial Research Promotion Fund (FFF no. 802746), Austrian Science Fund (FWF no. 8202) and Human Frontiers Science Program Organization.

Correspondence and requests for materials should be addressed to K.N. Supplementary Information is available on Nature Cell Biology’s World-Wide Web site (http://www.nature.com/ncb)

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  1. Research Institute of Molecular Pathology, Dr Bohr-Gasse 7, Vienna, A-1030, Austria

    Tomoyuki Tanaka & Kim Nasmyth

  2. Institute of Botany, University of Vienna, Rennweg 14, Vienna, A-1030, Austria

    Jörg Fuchs & Josef Loidl

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Supplementary information

Movie1

Sister centromeres split and reassociate repeatedly before moving finally to opposite spindle poles (Fig. 3b). (MOV 2145 kb)

Movie 2

Sister centromeres do not separate in an ndc 10-1 mutant at 37°C (Fig. 3c). (MOV 530 kb)

Movie 3

Sister centromeres split upon SPB duplication and do not reasso-ciate in the absence of Scc 1p (Fig. 4b). A cell of GAL-SCC 1, scc 1D strain. To deplete Scc 1p in the cells, they were grown in YEP-RaffGal, filtrated and cultured in YEP-Glc for 2 h before the time-lapse observation. (MOV 994 kb)

Movie 4

Sister centromeres split infrequently, and then only transiently, 4 soon after SPB duplication in the presence of Scc 1p (Fig. 4c; control of Movie 3 (Fig. 4b)). (MOV 1709 kb)

Movie 5

Both sister centromeres are frequently found moving in the vicin-ity of a single SPB in the absence of Scc 1p (Fig. 5a). A cell of GAL-SCC 1, scc 1D strain. Scc 1p was depleted as in Movie 3. (MOV 1338 kb)

The yeast cells shown in Fig.3b, c, 4b, c and 5a are displayed in movies 1-5. GFP and Nomarski images are presented. Both a locus at 1.4 kb from CEN 5 and spindle pole bodies (SPBs) were visualized as GFP dots. The speed of movies are 100 times faster than the real speed at which the events happen. All cells shown here harbour tetR-GFP, tetOs at 1.4 kb from CEN 5, SPC 42-GFP.

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Tanaka, T., Fuchs, J., Loidl, J. et al. Cohesin ensures bipolar attachment of microtubules to sister centromeres and resists their precocious separation. Nat Cell Biol 2, 492–499 (2000). https://doi.org/10.1038/35019529

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