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p27 binds cyclin–CDK complexes through a sequential mechanism involving binding-induced protein folding

Nature Structural & Molecular Biology volume 11, pages 358–364 (2004)Cite this article

Abstract

p27 controls cell proliferation by binding and regulating nuclear cyclin-dependent kinases (CDKs). In addition, p27 interacts with other nuclear and cytoplasmic targets and has diverse biological functions. We seek to understand how the structural and dynamic properties of p27 mediate its several functions. We show that, despite showing disorder before binding its targets, p27 has nascent secondary structure that may have a function in molecular recognition. Binding to Cdk2–cyclin A is accompanied by p27 folding, and kinetic data suggest a sequential mechanism that is initiated by binding to cyclin A. p27 regulates CDK–cyclin complexes involved directly in cell cycle control and does not interact with other closely related CDKs. We show that p27-cyclin interactions are an important determinant of this specificity and propose that the homologous cell cycle regulators p21 and p57 function by a similar sequential, folding-on-binding mechanism.

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Figure 1: Solution structure and dynamics of free p27-KID.
Figure 2: p27-KID binds Cdk2–cyclin A through a sequential mechanism involving extensive folding of p27-KID.
Figure 3: The molecular basis of p27 specificity for cell cycle CDKs.

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Acknowledgements

We thank W. Zhang and M. Stewart for assistance with NMR experiments, Y. Wang for assistance with protein production, C. Ross for computer support, P. Bowman for assistance with p27 structure figures and movie, G. Siuzdak, B. Bothner, C. Galea and T. Record for critically reading the manuscript and N. Pavletich for providing a cyclin A expression plasmid. R. Muhandiram and L. Kay are acknowledged for providing Varian NMR pulse sequences. The authors acknowledge support from the American Lebanese Syrian Associated Charities (ALSAC), the US National Cancer Institute (P30 CA 21765 and RO1 CA 82491), and the US National Center for Research Resources (S10 RR014675).

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  1. Igor Filippov

    Present address: Department of Chemistry, 1306 E. University Boulevard, Tucson, Arizona, 85721-0041, USA

Authors and Affiliations

  1. Department of Structural Biology, St. Jude Children's Research Hospital, 332, North Lauderdale Street, Memphis, 38105, Tennessee, USA

    Eilyn R Lacy, Igor Filippov, Steve Otieno, Limin Xiao & Richard W Kriwacki

  2. Hartwell Center, St. Jude Children's Research Hospital, 332, North Lauderdale Street, Memphis, 38105, Tennessee, USA

    William S Lewis

  3. Department of Molecular Sciences, University of Tennessee Health Sciences Center, Memphis, 38163, Tennessee, USA

    Steve Otieno

  4. Max-Planck-Institut für Biochemie, Am Klopferspitz 18a, Martinsried, D-82152, Germany

    Sonja Weiss & Ludger Hengst

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Correspondence to Richard W Kriwacki.

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Lacy, E., Filippov, I., Lewis, W. et al. p27 binds cyclin–CDK complexes through a sequential mechanism involving binding-induced protein folding. Nat Struct Mol Biol 11, 358–364 (2004). https://doi.org/10.1038/nsmb746

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