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Lookup NU author(s): Rhianna RowlandORCiD, Dr Richard Heath, Professor Jane Endicott, Professor Martin NobleORCiD, Dr Marco Salamina
This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0).
p27KIP1 (cyclin-dependent kinase inhibitor 1B, p27) is a member of the CIP/KIP family of CDK (cyclin dependent kinase) regulators that inhibit cell cycle CDKs. p27 phosphorylation by CDK1/2, signals its recruitment to the SCFSKP2 (S-phase kinase associated protein 1 (SKP1)-cullin-SKP2) E3 ubiquitin ligase complex for proteasomal degradation. The nature of p27 binding to SKP2 and CKS1 was revealed by the SKP1-SKP2-CKS1-p27 phosphopeptide crystal structure. Subsequently, a model for the hexameric CDK2-cyclin A-CKS1-p27-SKP1-SKP2 complex was proposed by overlaying an independently determined CDK2-cyclin A-p27 structure. Here we describe the experimentally determined structure of the isolated CDK2-cyclin A-CKS1-p27-SKP1-SKP2 complex at 3.4 Å global resolution using cryogenic electron microscopy. This structure supports previous analysis in which p27 was found to be structurally dynamic, transitioning from disordered to nascent secondary structure on target binding. We employed 3D variability analysis to further explore the conformational space of the hexameric complex and uncovered a previously unidentified hinge motion centred on CKS1. This flexibility gives rise to open and closed conformations of the hexameric complex that we propose may contribute to p27 regulation by facilitating recognition with SCFSKP2. This 3D variability analysis further informed particle subtraction and local refinement approaches to enhance the local resolution of the complex.
Author(s): Rowland RJ, Heath R, Maskell D, Thompson RF, Ranson NA, Blaza JN, Endicott JA, Noble MEM, Salamina M
Publication type: Article
Publication status: Published
Journal: Scientific Reports
Year: 2023
Volume: 13
Online publication date: 03/07/2023
Acceptance date: 24/06/2023
Date deposited: 20/06/2024
ISSN (electronic): 2045-2322
Publisher: Springer Nature
URL: https://doi.org/10.1038/s41598-023-37609-9
DOI: 10.1038/s41598-023-37609-9
Data Access Statement: The map and model of the hexameric CDK2-cyclin A-CKS1-p27-SKP1-SKP2 complex has been deposited in the Protein Data Bank and the Electron Microscopy Data Bank with accession numbers 8BYA, and 16325 respectively. The particle subtraction maps and models for SKP1-SKP2-CKS1 and CDK2-Cyclin A-p27 are deposited with accession codes PDB 8BYL/EMD-16327 and PDB 8BZO/EMD-16344 respectively.
PubMed id: 37400515
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