4NCI

Crystal Structure of Pyrococcus furiosis Rad50 R805E mutation

Summary for 4NCI

• Entry DOI: 10.2210/pdb4nci/pdb
• Related: 4NCH 4NCJ 4NCK
• Descriptor: DNA double-strand break repair Rad50 ATPase (2 entities in total)
• Functional Keywords: adenosine triphosphatase, dna repair, dna binding protein, fungal protein
• Biological source: Pyrococcus furiosus; More
• Total number of polymer chains: 1
• Total formula weight: 38580.54

Authors

Classen, S.,Williams, G.J.,Arvai, A.S.,Williams, R.S. (deposition date: 2013-10-24, release date: 2014-03-05, Last modification date: 2024-02-28)

Primary citation

Deshpande, R.A.,Williams, G.J.,Limbo, O.,Williams, R.S.,Kuhnlein, J.,Lee, J.H.,Classen, S.,Guenther, G.,Russell, P.,Tainer, J.A.,Paull, T.T.
ATP-driven Rad50 conformations regulate DNA tethering, end resection, and ATM checkpoint signaling.
Embo J., 33:482-500, 2014

PubMed Abstract: The Mre11-Rad50 complex is highly conserved, yet the mechanisms by which Rad50 ATP-driven states regulate the sensing, processing and signaling of DNA double-strand breaks are largely unknown. Here we design structure-based mutations in Pyrococcus furiosus Rad50 to alter protein core plasticity and residues undergoing ATP-driven movements within the catalytic domains. With this strategy we identify Rad50 separation-of-function mutants that either promote or destabilize the ATP-bound state. Crystal structures, X-ray scattering, biochemical assays, and functional analyses of mutant PfRad50 complexes show that the ATP-induced 'closed' conformation promotes DNA end binding and end tethering, while hydrolysis-induced opening is essential for DNA resection. Reducing the stability of the ATP-bound state impairs DNA repair and Tel1 (ATM) checkpoint signaling in Schizosaccharomyces pombe, double-strand break resection in Saccharomyces cerevisiae, and ATM activation by human Mre11-Rad50-Nbs1 in vitro, supporting the generality of the P. furiosus Rad50 structure-based mutational analyses. These collective results suggest that ATP-dependent Rad50 conformations switch the Mre11-Rad50 complex between DNA tethering, ATM signaling, and 5' strand resection, revealing molecular mechanisms regulating responses to DNA double-strand breaks.

PubMed: 24493214
DOI: 10.1002/embj.201386100

Experimental method

X-RAY DIFFRACTION (2.3 Å)