5DAC

ATP-gamma-S bound Rad50 from Chaetomium thermophilum in complex with DNA

Summary for 5DAC

• Entry DOI: 10.2210/pdb5dac/pdb
• Related: 5DA9
• Descriptor: Putative uncharacterized protein,Putative uncharacterized protein, DNA (5'-D(P*CP*CP*CP*CP*CP*CP*CP*CP*CP*CP*CP*CP*CP*CP*C)-3'), DNA (5'-D(P*GP*GP*GP*GP*GP*GP*GP*GP*GP*GP*GP*GP*GP*GP*G)-3'), ... (7 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: 111402.22

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: 2024-01-10)

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 (2.503 Å)