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Cooperative organization in a macromolecular complex

Nature Structural & Molecular Biology volume 10, pages 718–724 (2003)Cite this article

An Erratum to this article was published on 01 July 2004

Abstract

The mechanism of assembly of multiprotein complexes and the subsequent organization of activity are not well understood. Here we report the application of biophysical tools to investigate the relationship between structure and function in protein assemblies. We used as a model system the SCFSkp2 complex that targets p27Kip1 for ubiquitination and subsequent degradation; this process requires an adapter protein, Cks1. By dissecting the interactions between the different subunits we show that the properties of Cks1 are highly context dependent, and its activity is acquired only when the complex is fully assembled. The results provide insights into the central role of small adapters in macromolecular assembly and explain their high sequence conservation. Simultaneous and synergistic binding of multiple subunits in a complex provides the specificity and control required before the key cell-cycle regulator p27 is committed to degradation.

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Figure 1: Binding of fluorescein-labeled p27 phosphopeptide to Cks1.
Figure 2: Binding of Cks1 to Cdk2 (left) and Cks1 to Skp1-Skp2 (right).
Figure 3: Location of the three binding sites on Cks1 and effect of mutations in Cks1 on the affinity for p27 phosphopeptide.
Figure 4: Context-dependent signaling by the Cks1 adapter protein.

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Acknowledgements

We thank D. Vesprintsev for help with running the experiments, A. Hershko, M. Bycroft and S. Teichmann for helpful discussions, and B. Schulman and J. Endicott for providing clones. This work was supported by the Medical Research Council of the UK (MRC). L.S.I. was supported by a Career Development Award from the MRC, M.A.S. by an External Research Studentship from Trinity College, Cambridge, UK, and A.F. by a Human Frontier Science Program long-term fellowship.

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  1. Markus A Seeliger

    Present address: Howard Hughes Medical Institute, Department of Molecular and Cell Biology, Department of Chemistry, University of California, Berkeley, Berkeley, California, 94720, USA

  2. Sadie E Breward & Laura S Itzhaki

    Present address: MRC Cancer Cell Unit, Hutchison/MRC Research Centre, Hills Road, Cambridge, CB2 2XZ, UK

Authors and Affiliations

  1. MRC Centre for Protein Engineering, Hills Road, Cambridge, CB2 2QH, UK

    Markus A Seeliger, Sadie E Breward, Assaf Friedler, Oliver Schon & Laura S Itzhaki

  2. Cambridge University Chemical Laboratory, Lensfield Road, Cambridge, CB2 1EW, UK

    Markus A Seeliger, Sadie E Breward, Assaf Friedler, Oliver Schon & Laura S Itzhaki

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  1. Markus A Seeliger
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Correspondence to Laura S Itzhaki.

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Seeliger, M., Breward, S., Friedler, A. et al. Cooperative organization in a macromolecular complex. Nat Struct Mol Biol 10, 718–724 (2003). https://doi.org/10.1038/nsb962

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