PMID:20004108

Citation	Lage, C, Gonçalves, SR, Souza, LL, de Pádula, M and Leitão, AC (2010) Differential survival of Escherichia coli uvrA, uvrB, and uvrC mutants to psoralen plus UV-A (PUVA): Evidence for uncoupled action of nucleotide excision repair to process DNA adducts.J. Photochem. Photobiol. B, Biol. 98:40-7
Abstract	The nucleotide excision repair mechanism (NER) of Escherichia coli is responsible for the recognition and elimination of more than twenty different DNA lesions. Herein, we evaluated the in vivo role of NER in the repair of DNA adducts generated by psoralens (mono- or bi-functional) and UV-A light (PUVA) in E. coli. Cultures of wild-type E. coli K12 and mutants for uvrA, uvrB, uvrC or uvrAC genes were treated with PUVA and cell survival was determined. In parallel, kinetics of DNA repair was also evaluated by the comparison of DNA sedimentation profiles in all the strains after PUVA treatment. The uvrB mutant was more sensitive to PUVA treatment than all the other uvr mutant strains. Wild-type strain, and uvrA and uvrC mutants were able to repair PUVA-induced lesions, as seen by DNA sedimentation profiles, while the uvrB mutant was unable to repair the lesions. In addition, a quadruple fpg nth xth nfo mutant was unable to nick PUVA-treated DNA when the crude cell-free extract was used to perform plasmid nicking. These data suggest that DNA repair of PUVA-induced lesions may require base excision repair functions, despite proficient UvrABC activity. These results point to a specific role for UvrB protein in the repair of psoralen adducts, which appear to be independent of UvrA or UvrC proteins, as described for the classical UvrABC endonuclease mechanism.
Links	PubMed Online version:10.1016/j.jphotobiol.2009.11.001
Keywords	Adenosine Triphosphatases; DNA Adducts; DNA Helicases; DNA Repair; DNA-Binding Proteins; Endodeoxyribonucleases; Escherichia coli; Escherichia coli Proteins; Ficusin; Kinetics; Mutant Proteins; Photosensitizing Agents; Ultraviolet Rays
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Materials and Methods Used

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Phenotype Annotations

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Phenotype of	Taxon Information	Genotype Information (if known)	Condition Information	OMP ID	OMP Term Name	ECO ID	ECO Term Name	Notes	Status
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: AB1885 NCBI Taxon ID: 83333	Genotype of Reference Strain: uvrB+ Genotype of Experimental Strain : uvrB5	Reference Condition: UV-A + 8-MOP (0.1 microgram ml-1	OMP:0005098	decreased resistance to UV radiation	ECO:0000095	cell growth assay evidence	uvrB5 causes hypersensitivity to UV-A light treatments performed with bifunctional 8-methoxypsoralen when compared to wildtype, figure 1.
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: AB1886 NCBI Taxon ID: 83333	Genotype of Reference Strain: uvrA+ Genotype of Experimental Strain : uvrA6	Reference Condition: UV-A + 8-MOP (0.1 microgram ml-1	OMP:0005098	decreased resistance to UV radiation	ECO:0000095	cell growth assay evidence	uvrA6 causes increased sensitivity to UV-A light treatments performed with bifunctional 8-methoxypsoralen when compared to wildtype, figure 1.
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: AB1884 NCBI Taxon ID: 83333	Genotype of Reference Strain: uvrc+ Genotype of Experimental Strain : uvrC34	Reference Condition: UV-A + 8-MOP (0.1 microgram ml-1	OMP:0005098	decreased resistance to UV radiation	ECO:0000095	cell growth assay evidence	uvrC34 causes increased sensitivity to UV-A light treatments performed with bifunctional 8-methoxypsoralen when compared to wildtype, figure 1.
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: AB1885 NCBI Taxon ID: 83333	Genotype of Reference Strain: uvrB+ Genotype of Experimental Strain : uvrB5	Reference Condition: UV-A + 3-CPs (20 microgram ml-1	OMP:0005098	decreased resistance to UV radiation	ECO:0000095	cell growth assay evidence	uvrB5 causes hypersensitivity to UV-A light treatments performed with mono-functional 3-carbethoxypsoralen when compared to wildtype, figure 2.
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: AB1886 NCBI Taxon ID: 83333	Genotype of Reference Strain: uvrA+ Genotype of Experimental Strain : uvrA6	Reference Condition: UV-A + 3-CPs (20 microgram ml-1			ECO:0000095	cell growth assay evidence	uvrA6 causes unchanged sensitivity to UV-A light treatments performed with mono functional 3-carbethoxypsoralen when compared to wildtype, figure 2.
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: AB1884 NCBI Taxon ID: 83333	Genotype of Reference Strain: uvrC+ Genotype of Experimental Strain : uvrC34	Reference Condition: UV-A + 3-CPs (20 microgram ml-1			ECO:0000095	cell growth assay evidence	uvrC34 causes unchanged sensitivity to UV-A light treatments performed with mono functional 3-carbethoxypsoralen when compared to wildtype, figure 2.
a single strain under different conditions	Taxon: Escherichia coli Strain: K-12 Substrain: AB1157 NCBI Taxon ID: 83333	Genotype of Reference Strain: "uvrA+,uvrB+,uvrC+"	Reference Condition: Treatment with UV-A Experimental Condition: Treatment with UV-A + 8-MOP (0.1 microgram ml-1			ECO:0000095	cell growth assay evidence	The wildtype strain showed no change in UV-A sensitivity in the presence of bifunctional 8-methoxypsoralen.
a single strain under different conditions	Taxon: Escherichia coli Strain: K-12 Substrain: AB1157 NCBI Taxon ID: 83333	Genotype of Reference Strain: "uvrA+,uvrB+,uvrC+"	Reference Condition: Treatment with UV-A Experimental Condition: Treatment with UV-A + 3-CPs (20 microgram ml-1			ECO:0000095	cell growth assay evidence	The wildtype strain showed no change in UV-A sensitivity in the presence of monofunctional 3-carbethoxypsoralen.
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