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Structure of the Rad50 DNA double‐strand break repair protein in complex with DNA
Author(s) -
Rojowska Anna,
Lammens Katja,
Seifert Florian U,
Direnberger Carolin,
Feldmann Heidi,
Hopfner KarlPeter
Publication year - 2014
Publication title -
the embo journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 7.484
H-Index - 392
eISSN - 1460-2075
pISSN - 0261-4189
DOI - 10.15252/embj.201488889
Subject(s) - biology , rad50 , dna , dna repair , dna binding protein , replication protein a , genetics , dna clamp , microbiology and biotechnology , gene , rna , transcription factor , reverse transcriptase
The Mre11–Rad50 nuclease– ATP ase is an evolutionarily conserved multifunctional DNA double‐strand break ( DSB ) repair factor. Mre11–Rad50's mechanism in the processing, tethering, and signaling of DSB s is unclear, in part because we lack a structural framework for its interaction with DNA in different functional states. We determined the crystal structure of T hermotoga maritima Rad50 NBD (nucleotide‐binding domain) in complex with Mre11 HLH (helix‐loop‐helix domain), AMPPNP , and double‐stranded DNA . DNA binds between both coiled‐coil domains of the Rad50 dimer with main interactions to a strand‐loop‐helix motif on the NBD . Our analysis suggests that this motif on Rad50 does not directly recognize DNA ends and binds internal sites on DNA . Functional studies reveal that DNA binding to Rad50 is not critical for DNA double‐strand break repair but is important for telomere maintenance. In summary, we provide a structural framework for DNA binding to Rad50 in the ATP ‐bound state.