Structural Basis for the Activation and Target Site Specificity of CDC7 Kinase
| Autor: | Siobhan M. Hughes, Stefania Federico, Valerie E. Pye, Nicola O’Reilly, Samual D. Dick, Peter Cherepanov |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
DBF4
Cell Cycle Proteins Substrate Specificity INITIATION Structural Biology Catalytic Domain BINDING Phosphorylation 0303 health sciences biology Kinase Chemistry 030302 biochemistry & molecular biology Zinc Fingers Cell cycle DNA-DAMAGE CHECKPOINT Molecular Docking Simulation PHOSPHORYLATION SITES bisubstrate S-PHASE Target site cell cycle 03 Chemical Sciences Life Sciences & Biomedicine BISUBSTRATE INHIBITORS Protein Binding Biochemistry & Molecular Biology crystal structure kinase kinase inhibitor DDK Biophysics Protein Serine-Threonine Kinases Article CDC7 XL413 03 medical and health sciences zinc-binding domain Humans Molecular Biology S phase 030304 developmental biology Science & Technology Helicase Active site Cell Biology PROTEIN-KINASE 06 Biological Sciences MOLECULAR-MECHANISM Activation loop REPLICATION biology.protein CDC7-DBF4 08 Information and Computing Sciences |
| Zdroj: | 962.e4 Structure(London, England:1993) |
| ISSN: | 0969-2126 |
| DOI: | 10.1016/j.str.2020.05.010 |
| Popis: | Summary CDC7 is an essential Ser/Thr kinase that acts upon the replicative helicase throughout the S phase of the cell cycle and is activated by DBF4. Here, we present crystal structures of a highly active human CDC7-DBF4 construct. The structures reveal a zinc-finger domain at the end of the kinase insert 2 that pins the CDC7 activation loop to motif M of DBF4 and the C lobe of CDC7. These interactions lead to ordering of the substrate-binding platform and full opening of the kinase active site. In a co-crystal structure with a mimic of MCM2 Ser40 phosphorylation target, the invariant CDC7 residues Arg373 and Arg380 engage phospho-Ser41 at substrate P+1 position, explaining the selectivity of the S-phase kinase for Ser/Thr residues followed by a pre-phosphorylated or an acidic residue. Our results clarify the role of DBF4 in activation of CDC7 and elucidate the structural basis for recognition of its preferred substrates. Graphical Abstract Highlights • DBF4 activates CDC7 kinase via a two-step mechanism • Zinc-finger domain in CDC7 KI2 interacts with DBF4 motif M • Invariant CDC7 residues Arg373 and Arg380 engage P+1 substrate site CDC7 is a protein kinase that is essential for cell division. Using X-ray crystallography, Cherepanov and colleagues explain the two-step mechanism of CDC7 activation by its dedicated regulator protein DBF4. They also identify amino acid residues in CDC7 that are crucial for the recognition of its preferred substrates. |
| Databáze: | OpenAIRE |
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