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PDBsum entry 1w3s
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References listed in PDB file
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Key reference
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Title
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Crystal structure and DNA-Binding analysis of reco from deinococcus radiodurans.
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Authors
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I.Leiros,
J.Timmins,
D.R.Hall,
S.Mcsweeney.
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Ref.
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EMBO J, 2005,
24,
906-918.
[DOI no: ]
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PubMed id
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Abstract
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The RecFOR pathway has been shown to be essential for DNA repair through the
process of homologous recombination in bacteria and, recently, to be important
in the recovery of stalled replication forks following UV irradiation. RecO,
along with RecR, RecF, RecQ and RecJ, is a principal actor in this fundamental
DNA repair pathway. Here we present the three-dimensional structure of a member
of the RecO family. The crystal structure of Deinococcus radiodurans RecO
(drRecO) reveals possible binding sites for DNA and for the RecO-binding
proteins within its three discrete structural regions: an N-terminal
oligonucleotide/oligosaccharide-binding domain, a helical bundle and a
zinc-finger motif. Furthermore, drRecO was found to form a stable complex with
RecR and to bind both single- and double-stranded DNA. Mutational analysis
confirmed the existence of multiple DNA-binding sites within the protein.
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Figure 5.
Figure 5 Chromatogram of the gel filtration step for the RecOR
complex using a flow rate of 0.5 ml/min. The green and violet
lines show the absorbance at 280 and 260 nm, respectively. The
inset is the SDS -PAGE denaturing gel of the fractions as shown
above the chromatogram. Molecular weight markers (M) are shown
in kDa. drRecR (23.7 kDa) migrates as being slightly larger than
drRecO (26.3 kDa) and the proteins are at an apparent 2:1 ratio
in the RecOR complex.
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Figure 8.
Figure 8 Models for dsDNA interacting with drRecO based on the
DNA-binding studies and mutational analysis. In (A), the
secondary structure succession is outlined in colours ranging
from blue to red. Some positively charged residues are shown for
comparison to positive regions seen in the estimated
electrostatic surface potentials. Residues mutated in this study
are labelled in red. The electrostatic surface potentials in (B
-E) are contoured at  3
kT/e, where red describes a negative and blue a positive
potential. dsDNA interacting with drRecO is modelled as sticks
in (B, C). Two alternative binding sites involving the OB barrel
(bottom) and a positive patch (190-RHAVRRTVR-200) unique for
drRecO ending at the zinc-finger (top) are shown. (D) Close-up
of dsDNA modelled to interact with the positive patch unique to
drRecO with positively charged residues labelled. (E) Close-up
of the region in the OB barrel found to be important for dsDNA
binding in drRecO. (F) Indication of how well the mutants of
drRecO bind to DNA; +++, unaffected DNA-binding ability; +,
reduced DNA-binding ability; -, loss of DNA-binding ability.
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The above figures are
reprinted
from an Open Access publication published by Macmillan Publishers Ltd:
EMBO J
(2005,
24,
906-918)
copyright 2005.
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