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* Residue conservation analysis
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Gene Ontology (GO) functional annotation
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Cellular component
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intracellular
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1 term
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DOI no:
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Genes Dev
22:2034-2047
(2008)
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PubMed id:
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Structural and functional analysis of the Crb2-BRCT2 domain reveals distinct roles in checkpoint signaling and DNA damage repair.
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M.L.Kilkenny,
A.S.Doré,
S.M.Roe,
K.Nestoras,
J.C.Ho,
F.Z.Watts,
L.H.Pearl.
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ABSTRACT
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Schizosaccharomyces pombe Crb2 is a checkpoint mediator required for the
cellular response to DNA damage. Like human 53BP1 and Saccharomyces cerevisiae
Rad9 it contains Tudor(2) and BRCT(2) domains. Crb2-Tudor(2) domain interacts
with methylated H4K20 and is required for recruitment to DNA dsDNA breaks. The
BRCT(2) domain is required for dimerization, but its precise role in DNA damage
repair and checkpoint signaling is unclear. The crystal structure of the
Crb2-BRCT(2) domain, alone and in complex with a phosphorylated H2A.1 peptide,
reveals the structural basis for dimerization and direct interaction with
gamma-H2A.1 in ionizing radiation-induced foci (IRIF). Mutational analysis in
vitro confirms the functional role of key residues and allows the generation of
mutants in which dimerization and phosphopeptide binding are separately
disrupted. Phenotypic analysis of these in vivo reveals distinct roles in the
DNA damage response. Dimerization mutants are genotoxin sensitive and defective
in checkpoint signaling, Chk1 phosphorylation, and Crb2 IRIF formation, while
phosphopeptide-binding mutants are only slightly sensitive to IR, have extended
checkpoint delays, phosphorylate Chk1, and form Crb2 IRIF. However, disrupting
phosphopeptide binding slows formation of ssDNA-binding protein (Rpa1/Rad11)
foci and reduces levels of Rad22(Rad52) recombination foci, indicating a DNA
repair defect.
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Literature references that cite this PDB file's key reference
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PubMed id
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Reference
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M.Rappas,
A.W.Oliver,
and
L.H.Pearl
(2011).
Structure and function of the Rad9-binding region of the DNA-damage checkpoint adaptor TopBP1.
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Nucleic Acids Res, 39,
313-324.
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PDB codes:
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J.S.Williams,
R.S.Williams,
C.L.Dovey,
G.Guenther,
J.A.Tainer,
and
P.Russell
(2010).
gammaH2A binds Brc1 to maintain genome integrity during S-phase.
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EMBO J, 29,
1136-1148.
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PDB codes:
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M.Granata,
F.Lazzaro,
D.Novarina,
D.Panigada,
F.Puddu,
C.M.Abreu,
R.Kumar,
M.Grenon,
N.F.Lowndes,
P.Plevani,
and
M.Muzi-Falconi
(2010).
Dynamics of Rad9 chromatin binding and checkpoint function are mediated by its dimerization and are cell cycle-regulated by CDK1 activity.
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PLoS Genet, 6,
0.
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M.W.Richards,
J.W.Leung,
S.M.Roe,
K.Li,
J.Chen,
and
R.Bayliss
(2010).
A pocket on the surface of the N-terminal BRCT domain of Mcph1 is required to prevent abnormal chromosome condensation.
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J Mol Biol, 395,
908-915.
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PDB code:
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R.Cescutti,
S.Negrini,
M.Kohzaki,
and
T.D.Halazonetis
(2010).
TopBP1 functions with 53BP1 in the G1 DNA damage checkpoint.
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EMBO J, 29,
3723-3732.
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S.J.Campbell,
R.A.Edwards,
and
J.N.Glover
(2010).
Comparison of the structures and peptide binding specificities of the BRCT domains of MDC1 and BRCA1.
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Structure, 18,
167-176.
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PDB codes:
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S.L.Sanders,
A.R.Arida,
and
F.P.Phan
(2010).
Requirement for the phospho-H2AX binding module of Crb2 in double-strand break targeting and checkpoint activation.
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Mol Cell Biol, 30,
4722-4731.
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S.Pu,
A.L.Turinsky,
J.Vlasblom,
T.On,
X.Xiong,
A.Emili,
Z.Zhang,
J.Greenblatt,
J.Parkinson,
and
S.J.Wodak
(2010).
Expanding the landscape of chromatin modification (CM)-related functional domains and genes in human.
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PLoS One, 5,
e14122.
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S.Sofueva,
L.L.Du,
O.Limbo,
J.S.Williams,
and
P.Russell
(2010).
BRCT domain interactions with phospho-histone H2A target Crb2 to chromatin at double-strand breaks and maintain the DNA damage checkpoint.
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Mol Cell Biol, 30,
4732-4743.
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T.Ochi,
B.L.Sibanda,
Q.Wu,
D.Y.Chirgadze,
V.M.Bolanos-Garcia,
and
T.L.Blundell
(2010).
Structural biology of DNA repair: spatial organisation of the multicomponent complexes of nonhomologous end joining.
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J Nucleic Acids, 2010,
0.
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C.C.Nnakwe,
M.Altaf,
J.Côté,
and
S.J.Kron
(2009).
Dissection of Rad9 BRCT domain function in the mitotic checkpoint response to telomere uncapping.
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DNA Repair (Amst), 8,
1452-1461.
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T.Usui,
S.S.Foster,
and
J.H.Petrini
(2009).
Maintenance of the DNA-damage checkpoint requires DNA-damage-induced mediator protein oligomerization.
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Mol Cell, 33,
147-159.
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N.T.Greeson,
R.Sengupta,
A.R.Arida,
T.Jenuwein,
and
S.L.Sanders
(2008).
Di-methyl H4 lysine 20 targets the checkpoint protein Crb2 to sites of DNA damage.
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J Biol Chem, 283,
33168-33174.
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The most recent references are shown first.
Citation data come partly from CiteXplore and partly
from an automated harvesting procedure. Note that this is likely to be
only a partial list as not all journals are covered by
either method. However, we are continually building up the citation data
so more and more references will be included with time.
Where a reference describes a PDB structure, the PDB
codes are
shown on the right.
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Links
