Cryo-EM structure of SKP1-SKP2-CKS1 in complex with CDK2-cyclin A-p27KIP1
Abstract 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 u...
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| Format: | Article |
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Nature Portfolio
2023-07-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-023-37609-9 |
| _version_ | 1827905568041336832 |
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| author | Rhianna J. Rowland Richard Heath Daniel Maskell Rebecca F. Thompson Neil A. Ranson James N. Blaza Jane A. Endicott Martin E. M. Noble Marco Salamina |
| author_facet | Rhianna J. Rowland Richard Heath Daniel Maskell Rebecca F. Thompson Neil A. Ranson James N. Blaza Jane A. Endicott Martin E. M. Noble Marco Salamina |
| author_sort | Rhianna J. Rowland |
| collection | DOAJ |
| description | Abstract 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. |
| first_indexed | 2024-03-13T00:43:20Z |
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| id | doaj.art-255a782ee4174ccbb388f8fffc48d107 |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-03-13T00:43:20Z |
| publishDate | 2023-07-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-255a782ee4174ccbb388f8fffc48d1072023-07-09T11:10:34ZengNature PortfolioScientific Reports2045-23222023-07-0113111110.1038/s41598-023-37609-9Cryo-EM structure of SKP1-SKP2-CKS1 in complex with CDK2-cyclin A-p27KIP1Rhianna J. Rowland0Richard Heath1Daniel Maskell2Rebecca F. Thompson3Neil A. Ranson4James N. Blaza5Jane A. Endicott6Martin E. M. Noble7Marco Salamina8Translational and Clinical Research Institute, Newcastle University Centre for Cancer, Newcastle UniversityTranslational and Clinical Research Institute, Newcastle University Centre for Cancer, Newcastle UniversityAstbury Centre for Structural Molecular Biology, School of Molecular and Cellular Biology, University of LeedsAstbury Centre for Structural Molecular Biology, School of Molecular and Cellular Biology, University of LeedsAstbury Centre for Structural Molecular Biology, School of Molecular and Cellular Biology, University of LeedsDepartment of Chemistry, York Structural Biology Laboratory and York Biomedical Research Institute, University of YorkTranslational and Clinical Research Institute, Newcastle University Centre for Cancer, Newcastle UniversityTranslational and Clinical Research Institute, Newcastle University Centre for Cancer, Newcastle UniversityTranslational and Clinical Research Institute, Newcastle University Centre for Cancer, Newcastle UniversityAbstract 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.https://doi.org/10.1038/s41598-023-37609-9 |
| spellingShingle | Rhianna J. Rowland Richard Heath Daniel Maskell Rebecca F. Thompson Neil A. Ranson James N. Blaza Jane A. Endicott Martin E. M. Noble Marco Salamina Cryo-EM structure of SKP1-SKP2-CKS1 in complex with CDK2-cyclin A-p27KIP1 Scientific Reports |
| title | Cryo-EM structure of SKP1-SKP2-CKS1 in complex with CDK2-cyclin A-p27KIP1 |
| title_full | Cryo-EM structure of SKP1-SKP2-CKS1 in complex with CDK2-cyclin A-p27KIP1 |
| title_fullStr | Cryo-EM structure of SKP1-SKP2-CKS1 in complex with CDK2-cyclin A-p27KIP1 |
| title_full_unstemmed | Cryo-EM structure of SKP1-SKP2-CKS1 in complex with CDK2-cyclin A-p27KIP1 |
| title_short | Cryo-EM structure of SKP1-SKP2-CKS1 in complex with CDK2-cyclin A-p27KIP1 |
| title_sort | cryo em structure of skp1 skp2 cks1 in complex with cdk2 cyclin a p27kip1 |
| url | https://doi.org/10.1038/s41598-023-37609-9 |
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