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Degradation of a cohesin subunit by the N-end rule pathway is essential for chromosome stability

Nature volume 410, pages 955–959 (2001)Cite this article

Abstract

Cohesion between sister chromatids is established during DNA replication and depends on a protein complex called cohesin1,2,3,4,5,6,7. At the metaphase–anaphase transition in the yeast Saccharomyces cerevisiae, the ESP1-encoded protease separin cleaves SCC1, a subunit of cohesin with a relative molecular mass of 63,000 (Mr 63K)8. The resulting 33K carboxy-terminal fragment of SCC1 bears an amino-terminal arginine—a destabilizing residue in the N-end rule9. Here we show that the SCC1 fragment is short-lived (t1/2 ≈ 2 min), being degraded by the ubiquitin/proteasome-dependent N-end rule pathway. Overexpression of a long-lived derivative of the SCC1 fragment is lethal. In ubr1Δ cells, which lack the N-end rule pathway9, we found a highly increased frequency of chromosome loss. The bulk of increased chromosome loss in ubr1Δ cells is caused by metabolic stabilization of the ESP1-produced SCC1 fragment. This fragment is the first physiological substrate of the N-end rule pathway that is targeted through its N-terminal residue. A number of yeast proteins bear putative cleavage sites for the ESP1 separin, suggesting other physiological substrates and functions of the N-end rule pathway.

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Figure 1: The SCC1 fragment is a substrate of the N-end rule pathway.
Figure 2: Effects of overexpressing SCC1 and its fragments in UBR1 and ubr1Δ cells.
Figure 3: Increased chromosome instability in ubr1Δ S. cerevisiae.

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Acknowledgements

We are grateful to G. Turner, H.-R. Wang, F. Du, V. Ellison, A. Murray, D. Morgan, B. Stillman and P. Hieter for strains and plasmids. We thank M. Budd, W. Shuo and members of the Varshavsky laboratory, particularly F. Du and G. Turner, for helpful discussions. This work was supported by grants to A.V. from the NIH. H.R. is a Fellow of the Leukemia and Lymphoma Society. K.N. was supported by the Austrian Industrial Research Promotion Fund.

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Authors and Affiliations

  1. Division of Biology, California Institute of Technology, Pasadena, 91125, California, USA

    Hai Rao & Alexander Varshavsky

  2. Imperial Cancer Research Fund, 44 Lincoln's Inn Fields, London, WC2A 3PX, UK

    Frank Uhlmann

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

    Kim Nasmyth

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  1. Hai Rao
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Correspondence to Alexander Varshavsky.

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Rao, H., Uhlmann, F., Nasmyth, K. et al. Degradation of a cohesin subunit by the N-end rule pathway is essential for chromosome stability. Nature 410, 955–959 (2001). https://doi.org/10.1038/35073627

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