PDBsum entry 1w3s

DNA repair

PDB id: 1w3s

Contents

Protein chains

230 a.a. *

Metals

_ZN ×2

Waters ×23

* Residue conservation analysis

PDB id:

1w3s

Name:

DNA repair

Title:

The crystal structure of reco from deinococcus radiodurans.

Structure:

Hypothetical protein dr0819. Chain: a, b. Synonym: reco. Engineered: yes

Source:

Deinococcus radiodurans. Organism_taxid: 243230. Strain: r1. Expressed in: escherichia coli. Expression_system_taxid: 469008.

Resolution:

2.40Å R-factor: 0.227 R-free: 0.267

Authors:

I.Leiros,J.Timmins,D.R.Hall,G.A.Leonard,S.M.Mcsweeney

Key ref:

I.Leiros et al. (2005). Crystal structure and DNA-binding analysis of RecO from Deinococcus radiodurans. EMBO J, 24, 906-918. PubMed id: 15719017 DOI: 10.1038/sj.emboj.7600582

Date:

18-Jul-04 Release date: 23-Feb-05

Protein chains

Q9RW50  (Q9RW50_DEIRA) -  DNA repair protein RecO from Deinococcus radiodurans (strain ATCC 13939 / DSM 20539 / JCM 16871 / CCUG 27074 / LMG 4051 / NBRC 15346 / NCIMB 9279 / VKM B-1422 / R1)

Seq: 244 a.a.

Struc: 230 a.a.

Enzyme reactions

• Enzyme class: E.C.?

DOI no: 10.1038/sj.emboj.7600582

EMBO J 24:906-918 (2005)

PubMed id: 15719017

Crystal structure and DNA-binding analysis of RecO from Deinococcus radiodurans.

I.Leiros, J.Timmins, D.R.Hall, S.McSweeney.

ABSTRACT

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.

Selected figure(s)

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.

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.

The above figures are reprinted from an Open Access publication published by Macmillan Publishers Ltd: EMBO J (2005, 24, 906-918) copyright 2005.

Figures were selected by an automated process.