Reference: Deshpande I, et al. (2017) Structural Basis of Mec1-Ddc2-RPA Assembly and Activation on Single-Stranded DNA at Sites of Damage. Mol Cell 68(2):431-445.e5

Reference Help

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
Journal Article
Authors
Deshpande I, Seeber A, Shimada K, Keusch JJ, Gut H, Gasser SM
Primary Lit For
RFA1 | MEC1 | LCD1 | ATR-ATRIP DNA damage-sensing kinase complex

Interaction Annotations


Genetic Interactions

Increase the total number of rows showing on this page by using the pull-down located below the table, or use the page scroll at the table's top right to browse through the table's pages; use the arrows to the right of a column header to sort by that column; filter the table using the "Filter" box at the top of the table; click on the small "i" buttons located within a cell for an annotation to view further details about experiment type and any other genes involved in the interaction.

Interactor Interactor Allele Assay Annotation Action Phenotype SGA score P-value Source Reference

Physical Interactions 8 entries for 4 genes

Increase the total number of rows showing on this page by using the pull-down located below the table, or use the page scroll at the table's top right to browse through the table's pages; use the arrows to the right of a column header to sort by that column; filter the table using the "Filter" box at the top of the table; click on the small "i" buttons located within a cell for an annotation to view further details about experiment type and any other genes involved in the interaction.

InteractorInteractorAssayAnnotationActionModification
LCD1LCD1Co-crystal Structuremanually curatedHit-BaitNo Modification
LCD1LCD1Affinity Capture-Westernmanually curatedHit-BaitNo Modification
MEC1LCD1Affinity Capture-Westernmanually curatedHit-BaitNo Modification
RFA1LCD1Reconstituted Complexmanually curatedHit-BaitNo Modification
RFA1DNA2Protein-peptidemanually curatedHit-BaitNo Modification
RFA1LCD1Protein-peptidemanually curatedHit-BaitNo Modification
RFA1LCD1Co-crystal Structuremanually curatedHit-BaitNo Modification
RFA1LCD1Affinity Capture-Westernmanually curatedHit-BaitNo Modification
Showing 1 to 8 of 8 entries