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PDBsum entry 4nci
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DNA binding protein
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PDB id
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4nci
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References listed in PDB file
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Key reference
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Title
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Atp-Driven rad50 conformations regulate DNA tethering, End resection, And atm checkpoint signaling.
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Authors
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R.A.Deshpande,
G.J.Williams,
O.Limbo,
R.S.Williams,
J.Kuhnlein,
J.H.Lee,
S.Classen,
G.Guenther,
P.Russell,
J.A.Tainer,
T.T.Paull.
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Ref.
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Embo J, 2014,
33,
482-500.
[DOI no: ]
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PubMed id
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Abstract
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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.
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