The ASCC2 CUE domain in the ALKBH3–ASCC DNA repair complex recognizes adjacent ubiquitins in K63-linked polyubiquitin
| Autor: | Patrick M. Lombardi, Sara Haile, Timur Rusanov, Rebecca Rodell, Rita Anoh, Julia G. Baer, Kate A. Burke, Lauren N. Gray, Abigail R. Hacker, Kayla R. Kebreau, Christine K. Ngandu, Hannah A. Orland, Emmanuella Osei-Asante, Dhane P. Schmelyun, Devin E. Shorb, Shaheer H. Syed, Julianna M. Veilleux, Ananya Majumdar, Nima Mosammaparast, Cynthia Wolberger |
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| Rok vydání: | 2022 |
| Předmět: |
Models
Molecular Kd equilibrium dissociation constant DNA Repair nuclear magnetic resonance (NMR) DNA damage response ASCC2 Activating Signal Cointegrator 1 Complex Subunit 2 Biochemistry 03 medical and health sciences 0302 clinical medicine polyubiquitin ubiquitin ubiquitin-binding domain isothermal titration calorimetry (ITC) alkylation damage Ubiquitins Molecular Biology 030304 developmental biology 0303 health sciences ITC isothermal titration calorimetry Nuclear Proteins immunofluorescence microscopy ALKBH3 Alpha-ketoglutarate-dependent dioxygenase alkB homolog 3 DNA Cell Biology CSP chemical shift perturbation MMS methyl methanesulfonate TCEP Tris (2-carboxyethyl) phosphine CUE coupling of ubiquitin conjugation to ER degradation HADDOCK High Ambiguity Driven protein–protein DOCKing AlkB Homolog 3 Alpha-Ketoglutarate-Dependent Dioxygenase site-directed mutagenesis HSQC Heteronuclear Single Quantum Coherence signaling 030217 neurology & neurosurgery Research Article Protein Binding |
| Zdroj: | The Journal of Biological Chemistry |
| ISSN: | 0021-9258 |
| Popis: | Alkylation of DNA and RNA is a potentially toxic lesion that can result in mutations and even cell death. In response to alkylation damage, K63-linked polyubiquitin chains are assembled that localize the Alpha-ketoglutarate-dependent dioxygenase alkB homolog 3-Activating Signal Cointegrator 1 Complex Subunit (ASCC) repair complex to damage sites in the nucleus. The protein ASCC2, a subunit of the ASCC complex, selectively binds K63-linked polyubiquitin chains via its coupling of ubiquitin conjugation to ER degradation (CUE) domain. The basis for polyubiquitin-binding specificity was unclear, because CUE domains in other proteins typically bind a single ubiquitin and do not discriminate among different polyubiquitin linkage types. We report here that the ASCC2 CUE domain selectively binds K63-linked diubiquitin by contacting both the distal and proximal ubiquitin. The ASCC2 CUE domain binds the distal ubiquitin in a manner similar to that reported for other CUE domains bound to a single ubiquitin, whereas the contacts with the proximal ubiquitin are unique to ASCC2. Residues in the N-terminal portion of the ASCC2 α1 helix contribute to the binding interaction with the proximal ubiquitin of K63-linked diubiquitin. Mutation of residues within the N-terminal portion of the ASCC2 α1 helix decreases ASCC2 recruitment in response to DNA alkylation, supporting the functional significance of these interactions during the alkylation damage response. Our study reveals the versatility of CUE domains in ubiquitin recognition. |
| Databáze: | OpenAIRE |
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