8GNN

Crystal structure of the human RAD9-RAD1-HUS1-RAD17 complex

Summary for 8GNN

• Entry DOI: 10.2210/pdb8gnn/pdb
• Descriptor: Cell cycle checkpoint control protein RAD9A, Checkpoint protein HUS1, Cell cycle checkpoint protein RAD1, ... (5 entities in total)
• Functional Keywords: dna damage, checkpoint, dna repair, dna binding clamp, cell cycle
• Biological source: Homo sapiens (human); More
• Total number of polymer chains: 4
• Total formula weight: 95504.68

Authors

Hara, K.,Nagata, K.,Iida, N.,Hashimoto, H. (deposition date: 2022-08-24, release date: 2023-03-08, Last modification date: 2024-10-09)

Primary citation

Hara, K.,Hishiki, A.,Hoshino, T.,Nagata, K.,Iida, N.,Sawada, Y.,Ohashi, E.,Hashimoto, H.
The 9-1-1 DNA clamp subunit RAD1 forms specific interactions with clamp loader RAD17, revealing functional implications for binding-protein RHINO.
J.Biol.Chem., 299:103061-103061, 2023

PubMed Abstract: The RAD9-RAD1-HUS1 complex (9-1-1) is a eukaryotic DNA clamp with a crucial role at checkpoints for DNA damage. The ring-like structure of 9-1-1 is opened for loading onto 5' recessed DNA by the clamp loader RAD17 RFC-like complex (RAD17-RLC), in which the RAD17 subunit is responsible for specificity to 9-1-1. Loading of 9-1-1 is required for activation of the ATR-CHK1 checkpoint pathway and the activation is stimulated by a 9-1-1 interacting protein, RHINO, which interacts with 9-1-1 via a recently identified RAD1-binding motif. This discovery led to the hypothesis that other interacting proteins may contain a RAD1-binding motif as well. Here, we show that vertebrate RAD17 proteins also have a putative RAD1-binding motif in their N-terminal regions, and we report the crystal structure of human 9-1-1 bound to a human RAD17 peptide incorporating the motif at 2.1 Å resolution. Our structure confirms that the N-terminal region of RAD17 binds to the RAD1 subunit of 9-1-1 via specific interactions. Furthermore, we show that the RAD1-binding motif of RHINO disturbs the interaction of the N-terminal region of RAD17 with 9-1-1. Our results provide deeper understanding of how RAD17-RLC specifically recognizes 9-1-1 and imply that RHINO has a functional role in 9-1-1 loading/unloading and checkpoint activation.

PubMed: 36841485
DOI: 10.1016/j.jbc.2023.103061

Experimental method

X-RAY DIFFRACTION (2.119 Å)