5DA9

ATP-gamma-S bound Rad50 from Chaetomium thermophilum in complex with the Rad50-binding domain of Mre11

Summary for 5DA9

• Entry DOI: 10.2210/pdb5da9/pdb
• Descriptor: Putative uncharacterized protein,Putative uncharacterized protein, Putative double-strand break protein, MAGNESIUM ION, ... (5 entities in total)
• Functional Keywords: atpase, atpys bound, hydrolase
• Biological source: Chaetomium thermophilum var. thermophilum DSM 1495; More
• Total number of polymer chains: 4
• Total formula weight: 128183.29

Authors

Seifert, F.U.,Lammens, K.,Stoehr, G.,Kessler, B.,Hopfner, K.-P. (deposition date: 2015-08-19, release date: 2016-03-02, Last modification date: 2024-10-09)

Primary citation

Seifert, F.U.,Lammens, K.,Stoehr, G.,Kessler, B.,Hopfner, K.P.
Structural mechanism of ATP-dependent DNA binding and DNA end bridging by eukaryotic Rad50.
Embo J., 35:759-772, 2016

PubMed Abstract: The Mre11-Rad50-Nbs1 (MRN) complex is a central factor in the repair of DNA double-strand breaks (DSBs). The ATP-dependent mechanisms of how MRN detects and endonucleolytically processes DNA ends for the repair by microhomology-mediated end-joining or further resection in homologous recombination are still unclear. Here, we report the crystal structures of the ATPγS-bound dimer of the Rad50(NBD)(nucleotide-binding domain) from the thermophilic eukaryote Chaetomium thermophilum(Ct) in complex with either DNA or CtMre11(RBD)(Rad50-binding domain) along with small-angle X-ray scattering and cross-linking studies. The structure and DNA binding motifs were validated by DNA binding experiments in vitro and mutational analyses in Saccharomyces cerevisiae in vivo Our analyses provide a structural framework for the architecture of the eukaryotic Mre11-Rad50 complex. They show that a Rad50 dimer binds approximately 18 base pairs of DNA along the dimer interface in anATP-dependent fashion or bridges two DNA ends with a preference for 3' overhangs. Finally, our results may provide a general framework for the interaction of ABC ATPase domains of the Rad50/SMC/RecN protein family with DNA.

PubMed: 26896444
DOI: 10.15252/embj.201592934

Experimental method

X-RAY DIFFRACTION (3 Å)