Reference: Deshpande I, et al. (2017)
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Abstract
Mec1-Ddc2 (ATR-ATRIP) is a key DNA-damage-sensing kinase that is recruited through the single-stranded (ss) DNA-binding replication protein A (RPA) to initiate the DNA damage checkpoint response. Activation of ATR-ATRIP in the absence of DNA damage is lethal. Therefore, it is important that damage-specific recruitment precedes kinase activation, which is achieved at least in part by Mec1-Ddc2 homodimerization. Here, we report a structural, biochemical, and functional characterization of the yeast Mec1-Ddc2-RPA assembly. High-resolution co-crystal structures of Ddc2-Rfa1 and Ddc2-Rfa1-t11 (K45E mutant) N termini and of the Ddc2 coiled-coil domain (CCD) provide insight into Mec1-Ddc2 homodimerization and damage-site targeting. Based on our structural and functional findings, we present a Mec1-Ddc2-RPA-ssDNA composite structural model. By way of validation, we show that RPA-dependent recruitment of Mec1-Ddc2 is crucial for maintaining its homodimeric state at ssDNA and that Ddc2's recruitment domain and CCD are important for Mec1-dependent survival of UV-light-induced DNA damage.
- Reference Type
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Journal Article
- Authors
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Deshpande I,
Seeber A,
Shimada K,
Keusch JJ,
Gut H,
Gasser SM
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- RFA1 | MEC1 | LCD1 | ATR-ATRIP DNA damage-sensing kinase complex
Gene Ontology Annotations
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| Interactor | Interactor | Assay | Annotation | Action | Modification |
| LCD1 | LCD1 | Co-crystal Structure | manually curated | Hit-Bait | No Modification |
| LCD1 | LCD1 | Affinity Capture-Western | manually curated | Hit-Bait | No Modification |
| MEC1 | LCD1 | Affinity Capture-Western | manually curated | Hit-Bait | No Modification |
| RFA1 | LCD1 | Reconstituted Complex | manually curated | Hit-Bait | No Modification |
| RFA1 | DNA2 | Protein-peptide | manually curated | Hit-Bait | No Modification |
| RFA1 | LCD1 | Protein-peptide | manually curated | Hit-Bait | No Modification |
| RFA1 | LCD1 | Co-crystal Structure | manually curated | Hit-Bait | No Modification |
| RFA1 | LCD1 | Affinity Capture-Western | manually curated | Hit-Bait | No Modification |
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