PMID:20004108

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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

Main Points of the Paper

Please summarize the main points of the paper.

Materials and Methods Used

Please list the materials and methods used in this paper (strains, plasmids, antibodies, etc).

Phenotype Annotations

See Help:AnnotationTable for details on how to edit this table.

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:0007204

decreased resistance to UV-A radiation

ECO:0000095

cell growth assay evidence

Reference figure 1; 8-MOP=bifunctional 8-methoxypsoralen

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:0007204

decreased resistance to UV-A radiation

ECO:0000095

cell growth assay evidence

Reference figure 1; 8-MOP=bifunctional 8-methoxypsoralen

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:0007204

decreased resistance to UV-A radiation

ECO:0000095

cell growth assay evidence

Reference figure 1; 8-MOP=bifunctional 8-methoxypsoralen

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:0007204

decreased resistance to UV-A radiation

ECO:0000095

cell growth assay evidence

Reference figure 2; 3-CPs=mono-functional 3-carbethoxypsoralen

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)
OMP:0007198

UV-A radiation resistance

ECO:0000095

cell growth assay evidence

Reference figure 2; 3-CPs=mono-functional 3-carbethoxypsoralen; phenotype is unchanged

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)
OMP:0007198

UV-A radiation resistance

ECO:0000095

cell growth assay evidence

Reference figure 2; 3-CPs=mono-functional 3-carbethoxypsoralen; phenotype is unchanged

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)
OMP:0007198

UV-A radiation resistance

ECO:0000095

cell growth assay evidence

Inferred by curator from figure 1; 8-MOP=bifunctional 8-methoxypsoralen; phenotype is unchanged

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)
OMP:0007198

UV-A radiation resistance

ECO:0000095

cell growth assay evidence

Inferred by curator from figure 2; 3-CPs=monofunctional 3-carbethoxypsoralen; phenotype is unchanged

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-C treatment
OMP:0007201

decreased resistance to UV-C radiation

ECO:0000095

cell growth assay evidence

Reference figure 3

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-C treatment
OMP:0007201

decreased resistance to UV-C radiation

ECO:0000095

cell growth assay evidence

Reference figure 3

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-C treatment
OMP:0007201

decreased resistance to UV-C radiation

ECO:0000095

cell growth assay evidence

Reference figure 3

a mutation or genetic difference within a strain

  • Taxon: Escherichia coli
  • Strain: K-12
  • Substrain: JC3890
  • NCBI Taxon ID: 83333
  • Genotype of Reference Strain: uvrB+
  • Genotype of Experimental Strain : uvrB (del)
  • Reference Condition: UV-A + 8-MOP (.1 microgram ml-1)
OMP:0007204

decreased resistance to UV-A radiation

ECO:0000095

cell growth assay evidence

Reference figure 1; 8-MOP=bifunctional 8-methoxypsoralen

a mutation or genetic difference within a strain

  • Taxon: Escherichia coli
  • Strain: K-12
  • Substrain: JC3890
  • NCBI Taxon ID: 83333
  • Genotype of Reference Strain: uvrB+
  • Genotype of Experimental Strain : uvrB (del)
  • Reference Condition: UV-A + 3-CPs (20 microgram ml-1)
OMP:0007204

decreased resistance to UV-A radiation

ECO:0000095

cell growth assay evidence

Reference figure 2; 3-CPs=mono-functional 3-carbethoxypsoralen

a mutation or genetic difference within a strain

  • Taxon: Escherichia coli
  • Strain: K-12
  • Substrain: RJF604
  • NCBI Taxon ID: 83333
  • Genotype of Reference Strain: uvrC34 uvrA+
  • Genotype of Experimental Strain : uvrC34 uvrA::Tn10
  • Reference Condition: UV-A + 8-MOP (.1 microgram ml-1)
OMP:0007204

decreased resistance to UV-A radiation

ECO:0000095

cell growth assay evidence

Reference figure 1; 8-MOP=bifunctional 8-methoxypsoralen

a mutation or genetic difference within a strain

  • Taxon: Escherichia coli
  • Strain: K-12
  • Substrain: RJF604
  • NCBI Taxon ID: 83333
  • Genotype of Reference Strain: uvrC34 uvrA+
  • Genotype of Experimental Strain : uvrC34 uvrA::Tn10
  • Reference Condition: UV-A + 3-CPs (20 microgram ml-1)
OMP:0007198

UV-A radiation resistance

ECO:0000095

cell growth assay evidence

Reference figure 2; 3-CPs=monofunctional 3-carbethoxypsoralen; phenotype is unchanged

a mutation or genetic difference within a strain

  • Taxon: Escherichia coli
  • Strain: K-12
  • Substrain: RJF574
  • NCBI Taxon ID: 83333
  • Genotype of Reference Strain: uvrA+
  • Genotype of Experimental Strain : uvrA::Tn10
  • Reference Condition: UV-A + 8-MOP (.1 microgram ml-1)
OMP:0007204

decreased resistance to UV-A radiation

ECO:0000095

cell growth assay evidence

Reference figure 1; 8-MOP=bifunctional 8-methoxypsoralen

a mutation or genetic difference within a strain

  • Taxon: Escherichia coli
  • Strain: K-12
  • Substrain: RJF574
  • NCBI Taxon ID: 83333
  • Genotype of Reference Strain: uvrA+
  • Genotype of Experimental Strain : uvrA::Tn10
  • Reference Condition: UV-A + 3-CPs (20 microgram ml-1)
OMP:0007198

UV-A radiation resistance

ECO:0000095

cell growth assay evidence

Reference figure 2; 3-CPs=monofunctional 3-carbethoxypsoralen; phenotype is unchanged

a mutation or genetic difference within a strain

  • Taxon: Escherichia coli
  • Strain: K-12
  • Substrain: RJF573
  • NCBI Taxon ID: 83333
  • Genotype of Reference Strain: uvrC+
  • Genotype of Experimental Strain : uvrC::Tn10
  • Reference Condition: UV-A + 8-MOP (.1 microgram ml-1)
OMP:0007204

decreased resistance to UV-A radiation

ECO:0000095

cell growth assay evidence

Reference figure 1; 8-MOP=bifunctional 8-methoxypsoralen

a mutation or genetic difference within a strain

  • Taxon: Escherichia coli
  • Strain: K-12
  • Substrain: RJF573
  • NCBI Taxon ID: 83333
  • Genotype of Reference Strain: uvrC+
  • Genotype of Experimental Strain : uvrC::Tn10
  • Reference Condition: UV-A + 3-CPs (20 microgram ml-1)
OMP:0007198

UV-A radiation resistance

ECO:0000095

cell growth assay evidence

Reference figure 2; 3-CPs=monofunctional 3-carbethoxypsoralen; phenotype is unchanged

a mutation or genetic difference within a strain

  • Taxon: Escherichia coli
  • Strain: K-12
  • Substrain: JC3890
  • NCBI Taxon ID: 83333
  • Genotype of Reference Strain: uvrB+
  • Genotype of Experimental Strain : uvrB (del)
  • Reference Condition: UV-C treatment
OMP:0007201

decreased resistance to UV-C radiation

ECO:0000095

cell growth assay evidence

Reference figure 3

a mutation or genetic difference within a strain

  • Taxon: Escherichia coli
  • Strain: K-12
  • Substrain: RJF604
  • NCBI Taxon ID: 83333
  • Genotype of Reference Strain: uvrC34 uvrA+
  • Genotype of Experimental Strain : uvrC34 uvrA:Tn10
  • Reference Condition: UV-C treatment
OMP:0007201

decreased resistance to UV-C radiation

ECO:0000095

cell growth assay evidence

Reference figure 3

a mutation or genetic difference within a strain

  • Taxon: Escherichia coli
  • Strain: K-12
  • Substrain: RJF574
  • NCBI Taxon ID: 83333
  • Genotype of Reference Strain: uvrA+
  • Genotype of Experimental Strain : uvrA::Tn10
  • Reference Condition: UV-C treatment
OMP:0007201

decreased resistance to UV-C radiation

ECO:0000095

cell growth assay evidence

Reference figure 3

a mutation or genetic difference within a strain

  • Taxon: Escherichia coli
  • Strain: K-12
  • Substrain: RJF573
  • NCBI Taxon ID: 83333
  • Genotype of Reference Strain: uvrC+
  • Genotype of Experimental Strain : uvrC::Tn10
  • Reference Condition: UV-C treatment
OMP:0007201

decreased resistance to UV-C radiation

ECO:0000095

cell growth assay evidence

Reference figure 3

a mutation or genetic difference within a strain

  • Taxon: Escherichia coli
  • Strain: K-12
  • Substrain: RJF604
  • NCBI Taxon ID: 83333
  • Genotype of Reference Strain: uvrC34 uvrA+
  • Genotype of Experimental Strain : uvrC34 uvrA::Tn10
  • Reference Condition: UV-A + 8-MOP (0.1 microgram ml-1)
OMP:0007204

decreased resistance to UV-A radiation

ECO:0000095

cell growth assay evidence

Reference section 3.1; 8-MOP=bifunctional 8-methoxypsoralen

a mutation or genetic difference within a strain

  • Taxon: Escherichia coli
  • Strain: K-12
  • Substrain: RJF604
  • NCBI Taxon ID: 83333
  • Genotype of Reference Strain: uvrC34 uvrA+
  • Genotype of Experimental Strain : uvrC34 uvrA::Tn10
  • Reference Condition: UV-A + 3-CPs (20 microgram ml-1)
OMP:0007198

UV-A radiation resistance

ECO:0000095

cell growth assay evidence

Reference section 3.1; 3-CPs=monofunctional 3-carbethoxypsoralen

a mutation or genetic difference within a strain

  • Taxon: Escherichia coli
  • Strain: K-12
  • Substrain: RJF603
  • NCBI Taxon ID: 83333
  • Genotype of Reference Strain: uvrB (del) uvrA+
  • Genotype of Experimental Strain : uvrB (del) uvrA::Tn10
  • Reference Condition: UV-A + 8-MOP (0.1 microgram ml-1)
OMP:0007204

decreased resistance to UV-A radiation

ECO:0000095

cell growth assay evidence

Reference section 3.1; 8-MOP=bifunctional 8-methoxypsoralen

a mutation or genetic difference within a strain

  • Taxon: Escherichia coli
  • Strain: K-12
  • Substrain: RJF603
  • NCBI Taxon ID: 83333
  • Genotype of Reference Strain: uvrB (del) uvrA+
  • Genotype of Experimental Strain : uvrB (del) uvrA::Tn10
  • Reference Condition: UV-A + 3-CPs (20 microgram ml-1)
OMP:0007204

decreased resistance to UV-A radiation

ECO:0000095

cell growth assay evidence

Reference section 3.1; 3-CPs=monofunctional 3-carbethoxypsoralen

a mutation or genetic difference within a strain

  • Taxon: Escherichia coli
  • Strain: K-12
  • Substrain: RJF605
  • NCBI Taxon ID: 83333
  • Genotype of Reference Strain: uvrB (del) uvrC+
  • Genotype of Experimental Strain : uvrB (del) uvrC::Tn10
  • Reference Condition: UV-A + 8-MOP (0.1 microgram ml-1)
OMP:0007204

decreased resistance to UV-A radiation

ECO:0000095

cell growth assay evidence

Reference section 3.1; 8-MOP=bifunctional 8-methoxypsoralen

a mutation or genetic difference within a strain

  • Taxon: Escherichia coli
  • Strain: K-12
  • Substrain: RJF605
  • NCBI Taxon ID: 83333
  • Genotype of Reference Strain: uvrB (del) uvrC+
  • Genotype of Experimental Strain : uvrB (del) uvrC::Tn10
  • Reference Condition: UV-A + 3-CPs (20 microgram ml-1)
OMP:0007204

decreased resistance to UV-A radiation

ECO:0000095

cell growth assay evidence

Reference section 3.1; 3-CPs=monofunctional 3-carbethoxypsoralen



Notes

References

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