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The structural basis for specificity of substrate and recruitment peptides for cyclin-dependent kinases

Nature Cell Biology volume 1, pages 438–443 (1999)Cite this article

Abstract

Progression through the eukaryotic cell cycle is driven by the orderly activation of cyclin-dependent kinases (CDKs). For activity, CDKs require association with a cyclin and phosphorylation by a separate protein kinase at a conserved threonine residue (T160 in CDK2). Here we present the structure of a complex consisting of phosphorylated CDK2 and cyclin A together with an optimal peptide substrate, HHASPRK. This structure provides an explanation for the specificity of CDK2 towards the proline that follows the phosphorylatable serine of the substrate peptide, and the requirement for the basic residue in the P+3 position of the substrate. We also present the structure of phosphorylated CDK2 plus cyclin A3 in complex with residues 658–668 from the CDK2 substrate p107. These residues include the RXL motif required to target p107 to cyclins. This structure explains the specificity of the RXL motif for cyclins.

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Figure 1: The location of the peptide-binding sites on phospho-CDK2–cyclin A3 complex.
Figure 2: Contacts between CDK2 and the substrate peptide.
Figure 3: The recruitment-peptide-binding site of the phospho-CDK2–cyclin A3 complex.
Figure 4: The recruitment peptide inhibits the phosphorylation of GST–pRb (residues 792–928) but not phosphorylation of histone H1.

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Acknowledgements

We thank N. Hanlon for the GST–CDK2 GST–Cak1 coexpression vector; J. Hayles for the Schizosaccharomyces pombe p13suc1 peptide; J. Tucker for optimizing the phospho-CDK2 expression; and the beam-line scientists and staff at Elettra, Trieste, and ESRF station ID14 for their support during data collection. This work was supported by the Medical Research Council and the Royal Society.

Correspondence and requests for materials should be addressed to L.N.J. Coordinates have been deposited in Protein DataBank under accession number 1QMZ.

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  1. Nick R. Brown and Martin E. M. Noble: These authors contributed equally to this work

Authors and Affiliations

  1. Department of Biochemistry, Laboratory of Molecular Biophysics and Oxford Centre for Molecular Sciences, University of Oxford, Rex Richards Building, South Parks Road, Oxford, OX1 3QU, UK

    Nick R. Brown, Martin E. M. Noble, Jane A. Endicott & Louise N. Johnson

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  1. Nick R. Brown
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Correspondence to Louise N. Johnson.

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Brown, N., Noble, M., Endicott, J. et al. The structural basis for specificity of substrate and recruitment peptides for cyclin-dependent kinases. Nat Cell Biol 1, 438–443 (1999). https://doi.org/10.1038/15674

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