PMID:20727918

Citation	Prosser, GA, Patterson, AV and Ackerley, DF (2010) uvrB gene deletion enhances SOS chromotest sensitivity for nitroreductases that preferentially generate the 4-hydroxylamine metabolite of the anti-cancer prodrug CB1954.J. Biotechnol. 150:190-4
Abstract	CB1954 is an anti-cancer prodrug that can be reduced at either of two nitro groups to form cytotoxic metabolites. We describe here two efficient and previously uncharacterized nitroreductases, YfkO from Bacillus subtilis which reduces CB1954 exclusively at the 4-NO(2) position, and NfsA from Klebsiella pneumoniae which preferentially reduces the 2-NO(2) group. Utilizing these novel enzymes, together with three previously characterized nitroreductases, we demonstrate that the Escherichia coli SOS-chromotest assay can differentially detect the 4-nitro versus 2-nitro reduction products of CB1954 following deletion of the nucleotide excision repair gene uvrB, but not mismatch repair (mutS) or methyltransferase (ada/ogt) genes. These findings may hold significance for identification and selection of nitroreductases for CB1954-mediated gene therapy, particularly when targeting tumors that are deficient in nucleotide excision repair. Moreover, we demonstrate that comparative SOS chromotest analysis in wild type and uvrB mutant strains can be used to determine whether or not nucleotide excision repair plays a significant role in processing DNA damage resulting from activation of different nitroaromatic prodrugs.
Links	PubMed Online version:10.1016/j.jbiotec.2010.08.007
Keywords	Antineoplastic Agents; Aziridines; Bacillus subtilis; Bacterial Proteins; DNA Helicases; Escherichia coli Proteins; Gene Deletion; Kinetics; Klebsiella pneumoniae; Metronidazole; Molecular Conformation; Molecular Sequence Data; Nitrofurazone; Nitroreductases; SOS Response (Genetics)
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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: SOS-R1 NCBI Taxon ID: 83333	Genotype of Reference Strain: SOS-R1 pNfsA uvrB+ Genotype of Experimental Strain : SOS-R1 pNfsA uvrB(del)	Reference Condition: +CB1954 (50 micromolar for 3 hours) compared to 0 micromolar CB1954	OMP:0005023	presence of an SOS response	ECO:0000297	LacZ transcript localization evidence	Reference figure 2; NfsA (Ec)= 2-NO2 nitroreductase only, from E. coli; phenotype is unchanged. testing
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SOS-R1 NCBI Taxon ID: 83333	Genotype of Reference Strain: SOS-R1 pNfsA uvrB+ Genotype of Experimental Strain : SOS-R1 pNfsA uvrB(del)	Reference Condition: +CB1954 (50 micromolar for 3 hours) compared to 0 micromolar CB1954	OMP:0005023	presence of an SOS response	ECO:0000297	LacZ transcript localization evidence	Reference figure 2; NfsA (Kp)= 2-NO2 nitroreductase only, from K. pneumonia; phenotype is unchanged
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SOS-R1 NCBI Taxon ID: 83333	Genotype of Reference Strain: SOS-R1 pNfsB uvrB+ Genotype of Experimental Strain : SOS-R1 pNfsB uvrB(del)	Reference Condition: +CB1954 (50 micromolar for 3 hours) compared to 0 micromolar CB1954	OMP:0005023	presence of an SOS response	ECO:0000297	LacZ transcript localization evidence	Reference figure 2; NfsB= 2-NO2 or 4-NO2 nitroreductase; phenotype is unchanged
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SOS-R1 NCBI Taxon ID: 83333	Genotype of Reference Strain: SOS-R1 pNemA uvrB+ Genotype of Experimental Strain : SOS-R1 pNemA uvrB(del)	Reference Condition: +CB1954 (50 micromolar for 3 hours) compared to 0 micromolar CB1954	OMP:0005024	increased induced expression of the SOS response	ECO:0000297	LacZ transcript localization evidence	Reference figure 2; NemA= 4-NO2 nitroreductase only
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SOS-R1 NCBI Taxon ID: 83333	Genotype of Reference Strain: SOS-R1 pYfkO uvrB+ Genotype of Experimental Strain : SOS-R1 pYfkO uvrB(del)	Reference Condition: +CB1954 (50 micromolar for 3 hours) compared to 0 micromolar CB1954	OMP:0005024	increased induced expression of the SOS response	ECO:0000297	LacZ transcript localization evidence	Reference figure 2; Yfko (Bs)= 4-NO2 nitroreductase only, from B. licheniformis
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SOS-R1 NCBI Taxon ID: 83333	Genotype of Reference Strain: SOS-R1 pNfsA mutS+ Genotype of Experimental Strain : SOS-R1 pNfsA mutS(del)	Reference Condition: +CB1954 (50 micromolar for 3 hours) compared to 0 micromolar CB1954	OMP:0005023	presence of an SOS response	ECO:0000297	LacZ transcript localization evidence	Reference figure 2; NfsA (Ec)= 2-NO2 nitroreductase only, from E. coli; phenotype is unchanged
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SOS-R1 NCBI Taxon ID: 83333	Genotype of Reference Strain: SOS-R1 pNfsA mutS+ Genotype of Experimental Strain : SOS-R1 pNfsA mutS(del)	Reference Condition: +CB1954 (50 micromolar for 3 hours) compared to 0 micromolar CB1954	OMP:0005023	presence of an SOS response	ECO:0000297	LacZ transcript localization evidence	Reference figure 2; NfsA (Kp)= 2-NO2 nitroreductase only, from K. pneumoniae; phenotype is unchanged
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SOS-R1 NCBI Taxon ID: 83333	Genotype of Reference Strain: SOS-R1 pNfsB mutS+ Genotype of Experimental Strain : SOS-R1 pNfsB mutS(del)	Reference Condition: +CB1954 (50 micromolar for 3 hours) compared to 0 micromolar CB1954	OMP:0005023	presence of an SOS response	ECO:0000297	LacZ transcript localization evidence	Reference figure 2; NfsB= 2-NO2 or 4-NO2 nitroreductase; phenotype is unchanged
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SOS-R1 NCBI Taxon ID: 83333	Genotype of Reference Strain: SOS-R1 pNemA mutS+ Genotype of Experimental Strain : SOS-R1 pNemA mutS(del)	Reference Condition: +CB1954 (50 micromolar for 3 hours) compared to 0 micromolar CB1954	OMP:0005023	presence of an SOS response	ECO:0000297	LacZ transcript localization evidence	Reference figure 2; NemA= 4-NO2 nitroreductase only; phenotype is unchanged
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SOS-R1 NCBI Taxon ID: 83333	Genotype of Reference Strain: SOS-R1 pYfkO mutS+ Genotype of Experimental Strain : SOS-R1 pYfkO mutS(del)	Reference Condition: +CB1954 (50 micromolar for 3 hours) compared to 0 micromolar CB1954	OMP:0005023	presence of an SOS response	ECO:0000297	LacZ transcript localization evidence	Reference figure 2; Yfko (Bs)= 4-NO2 nitroreductase only, from B. licheniformis; phenotype is unchanged
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SOS-R1 NCBI Taxon ID: 83333	Genotype of Reference Strain: SOS-R1 pNfsA ada+ ogt+ Genotype of Experimental Strain : SOS-R1 pNfsA ada(del) ogt(del)	Reference Condition: +CB1954 (50 micromolar for 3 hours) compared to 0 micromolar CB1954	OMP:0005023	presence of an SOS response	ECO:0000297	LacZ transcript localization evidence	Reference figure 2; NfsA (Ec)= 2-NO2 nitroreductase only, from E. coli; phenotype is unchanged
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SOS-R1 NCBI Taxon ID: 83333	Genotype of Reference Strain: SOS-R1 NfsA uvrB+ mutS+ ada+ ogt+	Reference Condition: -CB1954 (0 micromolar for 3 hours) Experimental Condition: +CB1954 (50 micromolar for 3 hours)	OMP:0005024	increased induced expression of the SOS response	ECO:0000297	LacZ transcript localization evidence	Reference figure 2; NfsA (Ec)= 2-NO2 nitroreductase only, from E. coli
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SOS-R1 NCBI Taxon ID: 83333	Genotype of Reference Strain: SOS-R1 pNfsA ada+ ogt+ Genotype of Experimental Strain : SOS-R1 pNfsA ada(del) ogt(del)	Reference Condition: +CB1954 (50 micromolar for 3 hours) compared to 0 micromolar CB1954	OMP:0005023	presence of an SOS response	ECO:0000297	LacZ transcript localization evidence	Reference figure 2; NfsA (Kp)= 2-NO2 nitroreductase only, from K. pneumoniae; phenotype is unchanged
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SOS-R1 NCBI Taxon ID: 83333	Genotype of Reference Strain: SOS-R1 pNfsA uvrB+ mutS+ ada+ ogt+	Reference Condition: -CB1954 (0 micromolar for 3 hours) Experimental Condition: +CB1954 (50 micromolar for 3 hours)	OMP:0005024	increased induced expression of the SOS response	ECO:0000297	LacZ transcript localization evidence	Reference figure 2; NfsA (Kp)= 2-NO2 nitroreductase only, from K. pneumonia
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SOS-R1 NCBI Taxon ID: 83333	Genotype of Reference Strain: SOS-R1 pNfsB ada+ ogt+ Genotype of Experimental Strain : SOS-R1 pNfsB ada(del) ogt(del)	Reference Condition: +CB1954 (50 micromolar for 3 hours) compared to 0 micromolar CB1954	OMP:0005023	presence of an SOS response	ECO:0000297	LacZ transcript localization evidence	Reference figure 2; NfsB= 2-NO2 or 4-NO2 nitroreductase; phenotype is unchanged
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SOS-R1 NCBI Taxon ID: 83333	Genotype of Reference Strain: SOS-R1 pNfsB uvrB+ mutS+ ada+ ogt+	Reference Condition: -CB1954 (0 micromolar for 3 hours) Experimental Condition: +CB1954 (50 micromolar for 3 hours)	OMP:0005024	increased induced expression of the SOS response	ECO:0000297	LacZ transcript localization evidence	Reference figure 2; NfsB= 2-NO2 or 4-NO2 nitroreductase
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SOS-R1 NCBI Taxon ID: 83333	Genotype of Reference Strain: SOS-R1 pNemA ada+ ogt+ Genotype of Experimental Strain : SOS-R1 pNemA ada(del) ogt(del)	Reference Condition: +CB1954 (50 micromolar for 3 hours) compared to 0 micromolar CB1954	OMP:0005023	presence of an SOS response	ECO:0000297	LacZ transcript localization evidence	Reference figure 2; NemA= 4-NO2 nitroreductase only; phenotype is unchanged
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SOS-R1 NCBI Taxon ID: 83333	Genotype of Reference Strain: SOS-R1 NemA uvrB+ mutS+ ada+ ogt+	Reference Condition: -CB1954 (0 micromolar for 3 hours) Experimental Condition: +CB1954 (50 micromolar for 3 hours)	OMP:0005024	increased induced expression of the SOS response	ECO:0000297	LacZ transcript localization evidence	Reference figure 2; NemA= 4-NO2 nitroreductase only
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SOS-R1 NCBI Taxon ID: 83333	Genotype of Reference Strain: SOS-R1 pYfkO ada+ ogt+ Genotype of Experimental Strain : SOS-R1 pYfkO ada(del) ogt(del)	Reference Condition: +CB1954 (50 micromolar for 3 hours) compared to 0 micromolar CB1954	OMP:0005023	presence of an SOS response	ECO:0000297	LacZ transcript localization evidence	Reference figure 2; Yfko (Bs)= 4-NO2 nitroreductase only, from B. licheniformis; phenotype is unchanged
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SOS-R1 NCBI Taxon ID: 83333	Genotype of Reference Strain: SOS-R1 YfkO uvrB+ mutS+ ada+ ogt+	Reference Condition: -CB1954 (0 micromolar for 3 hours) Experimental Condition: +CB1954 (50 micromolar for 3 hours)	OMP:0005024	increased induced expression of the SOS response	ECO:0000297	LacZ transcript localization evidence	Reference figure 2; Yfko (Bs)= 4-NO2 nitroreductase only, from B. licheniformis
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SOS-R1 NCBI Taxon ID: 83333	Genotype of Reference Strain: SOS-R1 NfsA uvrB(del)	Reference Condition: -CB1954 (0 micromolar for 3 hours) Experimental Condition: +CB1954 (50 micromolar for 3 hours)	OMP:0005024	increased induced expression of the SOS response	ECO:0000297	LacZ transcript localization evidence	Reference figure 2; NfsA (Ec)= 2-NO2 nitroreductase only, from E. coli test
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SOS-R1 83333	Genotype of Reference Strain: SOS-R1 NfsA uvrB(del)	Reference Condition: -CB1954 (0 micromolar for 3 hours) Experimental Condition: +CB1954 (50 micromolar for 3 hours)	OMP:0005024	increased induced expression of the SOS response	ECO:0000297	LacZ transcript localization evidence	Reference figure 2; NfsA (Kp)= 2-NO2 nitroreductase only, from K. pneumonia
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SOS-R1 83333	Genotype of Reference Strain: SOS-R1 NfsB uvrB(del)	Reference Condition: -CB1954 (0 micromolar for 3 hours) Experimental Condition: +CB1954 (50 micromolar for 3 hours)	OMP:0005024	increased induced expression of the SOS response	ECO:0000297	LacZ transcript localization evidence	Reference figure 2; NfsB= 2-NO2 or 4-NO2 nitroreductase
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SOS-R1 83333	Genotype of Reference Strain: SOS-R1 NemA uvrB(del)	Reference Condition: -CB1954 (0 micromolar for 3 hours) Experimental Condition: +CB1954 (50 micromolar for 3 hours)	OMP:0005024	increased induced expression of the SOS response	ECO:0000297	LacZ transcript localization evidence	Reference figure 2; NemA= 4-NO2 nitroreductase only
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SOS-R1 83333	Genotype of Reference Strain: SOS-R1 YfkO uvrB(del)	Reference Condition: -CB1954 (0 micromolar for 3 hours) Experimental Condition: +CB1954 (50 micromolar for 3 hours)	OMP:0005024	increased induced expression of the SOS response	ECO:0000297	LacZ transcript localization evidence	Reference figure 2; Yfko (Bs)= 4-NO2 nitroreductase only, from B. licheniformis
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SOS-R1 83333	Genotype of Reference Strain: SOS-R1 NfsA mutS(del)	Reference Condition: -CB1954 (0 micromolar for 3 hours) Experimental Condition: +CB1954 (50 micromolar for 3 hours)	OMP:0005024	increased induced expression of the SOS response	ECO:0000297	LacZ transcript localization evidence	Reference figure 2; NfsA (Ec)= 2-NO2 nitroreductase only, from E. coli
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SOS-R1 83333	Genotype of Reference Strain: SOS-R1 NfsA mutS(del)	Reference Condition: -CB1954 (0 micromolar for 3 hours) Experimental Condition: +CB1954 (50 micromolar for 3 hours)	OMP:0005024	increased induced expression of the SOS response	ECO:0000297	LacZ transcript localization evidence	Reference figure 2; NfsA (Kp)= 2-NO2 nitroreductase only, from K. pneumonia
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SOS-R1 83333	Genotype of Reference Strain: SOS-R1 NfsB mutS(del)	Reference Condition: -CB1954 (0 micromolar for 3 hours) Experimental Condition: +CB1954 (50 micromolar for 3 hours)	OMP:0005024	increased induced expression of the SOS response	ECO:0000297	LacZ transcript localization evidence	Reference figure 2; NfsB= 2-NO2 or 4-NO2 nitroreductase
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SOS-R1 83333	Genotype of Reference Strain: SOS-R1 NemA mutS(del)	Reference Condition: -CB1954 (0 micromolar for 3 hours) Experimental Condition: +CB1954 (50 micromolar for 3 hours)	OMP:0005024	increased induced expression of the SOS response	ECO:0000297	LacZ transcript localization evidence	Reference figure 2; NemA= 4-NO2 nitroreductase only
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SOS-R1 83333	Genotype of Reference Strain: SOS-R1 YfkO mutS(del)	Reference Condition: -CB1954 (0 micromolar for 3 hours) Experimental Condition: +CB1954 (50 micromolar for 3 hours)	OMP:0005024	increased induced expression of the SOS response	ECO:0000297	LacZ transcript localization evidence	Reference figure 2; Yfko (Bs)= 4-NO2 nitroreductase only, from B. licheniformis
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SOS-R1 83333	Genotype of Reference Strain: SOS-R1 NfsA ada(del) ogt(del)	Reference Condition: -CB1954 (0 micromolar for 3 hours) Experimental Condition: +CB1954 (50 micromolar for 3 hours)	OMP:0005024	increased induced expression of the SOS response	ECO:0000297	LacZ transcript localization evidence	Reference figure 2; NfsA (Ec)= 2-NO2 nitroreductase only, from E. coli
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SOS-R1 83333	Genotype of Reference Strain: SOS-R1 NfsA ada(del) ogt(del)	Reference Condition: -CB1954 (0 micromolar for 3 hours) Experimental Condition: +CB1954 (50 micromolar for 3 hours)	OMP:0005024	increased induced expression of the SOS response	ECO:0000297	LacZ transcript localization evidence	Reference figure 2; NfsA (Kp)= 2-NO2 nitroreductase only, from K. pneumonia
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SOS-R1 83333	Genotype of Reference Strain: SOS-R1 NfsB ada(del) ogt(del)	Reference Condition: -CB1954 (0 micromolar for 3 hours) Experimental Condition: +CB1954 (50 micromolar for 3 hours)	OMP:0005024	increased induced expression of the SOS response	ECO:0000297	LacZ transcript localization evidence	Reference figure 2; NfsB= 2-NO2 or 4-NO2 nitroreductase
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SOS-R1 83333	Genotype of Reference Strain: SOS-R1 NemA ada(del) ogt(del)	Reference Condition: -CB1954 (0 micromolar for 3 hours) Experimental Condition: +CB1954 (50 micromolar for 3 hours)	OMP:0005024	increased induced expression of the SOS response	ECO:0000297	LacZ transcript localization evidence	Reference figure 2; NemA= 4-NO2 nitroreductase only
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SOS-R1 83333	Genotype of Reference Strain: SOS-R1 YfkO ada(del) ogt(del)	Reference Condition: -CB1954 (0 micromolar for 3 hours) Experimental Condition: +CB1954 (50 micromolar for 3 hours)	OMP:0005024	increased induced expression of the SOS response	ECO:0000297	LacZ transcript localization evidence	Reference figure 2; Yfko (Bs)= 4-NO2 nitroreductase only, from B. licheniformis
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SOS-R1 83333	Genotype of Reference Strain: SOS-R1 NfsA uvrB+ Genotype of Experimental Strain: SOS-R1 NfsA uvrB(del)	Reference Condition: 5-nitrofuran nitrofurazone (5 micromolar for 3 hours compared to 0 micromolar)	OMP:0005024	increased induced expression of the SOS response	ECO:0000297	LacZ transcript localization evidence	Reference figure 3; NfsA (Ec)= 2-NO2 nitroreductase only, from E. coli
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SOS-R1 83333	Genotype of Reference Strain: SOS-R1 NfsA uvrB+ Genotype of Experimental Strain: SOS-R1 NfsA uvrB(del)	Reference Condition: 5-nitrofuran nitrofurazone (5 micromolar for 3 hours compared to 0 micromolar)	OMP:0005024	increased induced expression of the SOS response	ECO:0000297	LacZ transcript localization evidence	Reference figure 3; NfsA (Kp)= 2-NO2 nitroreductase only, from K. pneumonia
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SOS-R1 83333	Genotype of Reference Strain: SOS-R1 NfsB uvrB+ Genotype of Experimental Strain: SOS-R1 NfsB uvrB(del)	Reference Condition: 5-nitrofuran nitrofurazone (5 micromolar for 3 hours compared to 0 micromolar)	OMP:0005024	increased induced expression of the SOS response	ECO:0000297	LacZ transcript localization evidence	Reference figure 3; NfsB= 2-NO2 or 4-NO2 nitroreductase
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SOS-R1 83333	Genotype of Reference Strain: SOS-R1 NemA uvrB+ Genotype of Experimental Strain: SOS-R1 NemA uvrB(del)	Reference Condition: 5-nitrofuran nitrofurazone (5 micromolar for 3 hours compared to 0 micromolar)	OMP:0005024	increased induced expression of the SOS response	ECO:0000297	LacZ transcript localization evidence	Reference figure 3; NemA= 4-NO2 nitroreductase only
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SOS-R1 83333	Genotype of Reference Strain: SOS-R1 YfkO uvrB+ Genotype of Experimental Strain: SOS-R1 YfkO uvrB(del)	Reference Condition: 5-nitrofuran nitrofurazone (5 micromolar for 3 hours compared to 0 micromolar)	OMP:0005024	increased induced expression of the SOS response	ECO:0000297	LacZ transcript localization evidence	Reference figure 3; Yfko (Bs)= 4-NO2 nitroreductase only, from B. licheniformis
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SOS-R1 83333	Genotype of Reference Strain: SOS-R1 NfsA uvrB+ Genotype of Experimental Strain: SOS-R1 NfsA uvrB(del)	Reference Condition: 5-nitroimidazole metronidazole (5 micromolar for 3 hours compared to 0 micromolar)	OMP:0005023	presence of an SOS response	ECO:0000297	LacZ transcript localization evidence	Reference figure 3; NfsA (Ec)= 2-NO2 nitroreductase only, from E. coli; phenotype is unchanged
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SOS-R1 83333	Genotype of Reference Strain: SOS-R1 NfsA uvrB+ Genotype of Experimental Strain: SOS-R1 NfsA uvrB(del)	Reference Condition: 5-nitroimidazole metronidazole (5 micromolar for 3 hours compared to 0 micromolar)	OMP:0005023	presence of an SOS response	ECO:0000297	LacZ transcript localization evidence	Reference figure 3; NfsA (Kp)= 2-NO2 nitroreductase only, from K. pneumonia; phenotype is unchanged
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SOS-R1 83333	Genotype of Reference Strain: SOS-R1 NfsB uvrB+ Genotype of Experimental Strain: SOS-R1 NfsB uvrB(del)	Reference Condition: 5-nitroimidazole metronidazole (5 micromolar for 3 hours compared to 0 micromolar)	OMP:0005023	presence of an SOS response	ECO:0000297	LacZ transcript localization evidence	Reference figure 3; NfsB= 2-NO2 or 4-NO2 nitroreductase; phenotype is unchanged
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SOS-R1 83333	Genotype of Reference Strain: SOS-R1 NemA uvrB+ Genotype of Experimental Strain: SOS-R1 NemA uvrB(del)	Reference Condition: 5-nitroimidazole metronidazole (5 micromolar for 3 hours compared to 0 micromolar)	OMP:0005023	presence of an SOS response	ECO:0000297	LacZ transcript localization evidence	Reference figure 3; NemA= 4-NO2 nitroreductase only; phenotype is unchanged
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SOS-R1 83333	Genotype of Reference Strain: SOS-R1 YfkO uvrB+ Genotype of Experimental Strain: SOS-R1 YfkO uvrB(del)	Reference Condition: 5-nitroimidazole metronidazole (5 micromolar for 3 hours compared to 0 micromolar)	OMP:0005023	presence of an SOS response	ECO:0000297	LacZ transcript localization evidence	Reference figure 3; Yfko (Bs)= 4-NO2 nitroreductase only, from B. licheniformis; phenotype is unchanged
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SOS-R1 83333	Genotype of Reference Strain: SOS-R1 NfsA mutS+ Genotype of Experimental Strain: SOS-R1 NfsA mutS(del)	Reference Condition: 5-nitroimidazole metronidazole (5 micromolar for 3 hours compared to 0 micromolar)	OMP:0005023	presence of an SOS response	ECO:0000297	LacZ transcript localization evidence	Reference section 3.3; NfsA (Ec)= 2-NO2 nitroreductase only, from E. coli; phenotype is unchanged
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SOS-R1 83333	Genotype of Reference Strain: SOS-R1 NfsA mutS+ Genotype of Experimental Strain: SOS-R1 NfsA mutS(del)	Reference Condition: 5-nitroimidazole metronidazole (5 micromolar for 3 hours compared to 0 micromolar)	OMP:0005023	presence of an SOS response	ECO:0000297	LacZ transcript localization evidence	Reference section 3.3; NfsA (Kp)= 2-NO2 nitroreductase only, from K. pneumonia; phenotype is unchanged
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SOS-R1 83333	Genotype of Reference Strain: SOS-R1 NfsB mutS+ Genotype of Experimental Strain: SOS-R1 NfsB mutS(del)	Reference Condition: 5-nitroimidazole metronidazole (5 micromolar for 3 hours compared to 0 micromolar)	OMP:0005023	presence of an SOS response	ECO:0000297	LacZ transcript localization evidence	Reference section 3.3; NfsB= 2-NO2 or 4-NO2 nitroreductase; phenotype is unchanged
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SOS-R1 83333	Genotype of Reference Strain: SOS-R1 NemA mutS+ Genotype of Experimental Strain: SOS-R1 NemA mutS(del)	Reference Condition: 5-nitroimidazole metronidazole (5 micromolar for 3 hours compared to 0 micromolar)	OMP:0005023	presence of an SOS response	ECO:0000297	LacZ transcript localization evidence	Reference section 3.3; NemA= 4-NO2 nitroreductase only; phenotype is unchanged
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SOS-R1 83333	Genotype of Reference Strain: SOS-R1 YfkO mutS+ Genotype of Experimental Strain: SOS-R1 YfkO mutS(del)	Reference Condition: 5-nitroimidazole metronidazole (5 micromolar for 3 hours compared to 0 micromolar)	OMP:0005023	presence of an SOS response	ECO:0000297	LacZ transcript localization evidence	Reference section 3.3; YfkO (Bs)= 4-NO2 nitroreductase only, from B. licheniformis; phenotype is unchanged
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SOS-R1 83333	Genotype of Reference Strain: SOS-R1 NfsA mutS+ Genotype of Experimental Strain: SOS-R1 NfsA mutS(del)	Reference Condition: 5-nitrofuran nitrofurazone(5 micromolar for 3 hours compared to 0 micromolar)	OMP:0005023	presence of an SOS response	ECO:0000297	LacZ transcript localization evidence	Reference section 3.3; NfsA (Ec)= 2-NO2 nitroreductase only, from E. coli; phenotype is unchanged
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SOS-R1 83333	Genotype of Reference Strain: SOS-R1 NfsA mutS+ Genotype of Experimental Strain: SOS-R1 NfsA mutS(del)	Reference Condition: 5-nitrofuran nitrofurazone(5 micromolar for 3 hours compared to 0 micromolar)	OMP:0005023	presence of an SOS response	ECO:0000297	LacZ transcript localization evidence	Reference section 3.3; NfsA (Kp)= 2-NO2 nitroreductase only, from K. pneumonia; phenotype is unchanged
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SOS-R1 83333	Genotype of Reference Strain: SOS-R1 NfsB mutS+ Genotype of Experimental Strain: SOS-R1 NfsB mutS(del)	Reference Condition: 5-nitrofuran nitrofurazone(5 micromolar for 3 hours compared to 0 micromolar)	OMP:0005023	presence of an SOS response	ECO:0000297	LacZ transcript localization evidence	Reference section 3.3; NfsB= 2-NO2 or 4-NO2 nitroreductase; phenotype is unchanged
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SOS-R1 83333	Genotype of Reference Strain: SOS-R1 NemA mutS+ Genotype of Experimental Strain: SOS-R1 NemA mutS(del)	Reference Condition: 5-nitrofuran nitrofurazone(5 micromolar for 3 hours compared to 0 micromolar)	OMP:0005023	presence of an SOS response	ECO:0000297	LacZ transcript localization evidence	Reference section 3.3; NemA= 4-NO2 nitroreductase only; phenotype is unchanged
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SOS-R1 83333	Genotype of Reference Strain: SOS-R1 YfkO mutS+ Genotype of Experimental Strain: SOS-R1 YfkO mutS(del)	Reference Condition: 5-nitrofuran nitrofurazone(5 micromolar for 3 hours compared to 0 micromolar)	OMP:0005023	presence of an SOS response	ECO:0000297	LacZ transcript localization evidence	Reference section 3.3; YfkO (Bs)= 4-NO2 nitroreductase only, from B. licheniformis; phenotype is unchanged xx
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SOS-R1 NCBI Taxon ID: 83333	Genotype of Reference Strain: SOS-R1 NfsA ada+ ogt+ Genotype of Experimental Strain : SOS-R1 NfsA ada(del) ogt(del)	Reference Condition: 5-nitroimidazole metronidazole (5 micromolar for 3 hours compared to 0 micromolar)	OMP:0005023	presence of an SOS response	ECO:0000297	LacZ transcript localization evidence	Reference section 3.3; NfsA (Ec)= 2-NO2 nitroreductase only, from E. coli; phenotype is unchanged; phenotype is unchanged yy
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SOS-R1 83333	Genotype of Reference Strain: SOS-R1 NfsA ada+ ogt+ Genotype of Experimental Strain: SOS-R1 NfsA ada(del) ogt(del)	Reference Condition: 5-nitroimidazole metronidazole (5 micromolar for 3 hours compared to 0 micromolar)	OMP:0005023	presence of an SOS response	ECO:0000297	LacZ transcript localization evidence	Reference section 3.3; NfsA (Kp)= 2-NO2 nitroreductase only, from K. pneumonia; phenotype is unchanged
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SOS-R1 83333	Genotype of Reference Strain: SOS-R1 NfsB ada+ ogt+ Genotype of Experimental Strain: SOS-R1 NfsB ada(del) ogt(del)	Reference Condition: 5-nitroimidazole metronidazole (5 micromolar for 3 hours compared to 0 micromolar)	OMP:0005023	presence of an SOS response	ECO:0000297	LacZ transcript localization evidence	Reference section 3.3; NfsB= 2-NO2 or 4-NO2 nitroreductase; phenotype is unchanged
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SOS-R1 83333	Genotype of Reference Strain: SOS-R1 NemA ada+ ogt+ Genotype of Experimental Strain: SOS-R1 NemA ada(del) ogt(del)	Reference Condition: 5-nitroimidazole metronidazole (5 micromolar for 3 hours compared to 0 micromolar)	OMP:0005023	presence of an SOS response	ECO:0000297	LacZ transcript localization evidence	Reference section 3.3; NemA= 4-NO2 nitroreductase only; phenotype is unchanged
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SOS-R1 83333	Genotype of Reference Strain: SOS-R1 YfkO ada+ ogt+ Genotype of Experimental Strain: SOS-R1 YfkO ada(del) ogt(del)	Reference Condition: 5-nitroimidazole metronidazole (5 micromolar for 3 hours compared to 0 micromolar)	OMP:0005023	presence of an SOS response	ECO:0000297	LacZ transcript localization evidence	Reference section 3.3; YfkO (Bs)= 4-NO2 nitroreductase only, from B. licheniformis; phenotype is unchanged
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SOS-R1 83333	Genotype of Reference Strain: SOS-R1 NfsA ada+ ogt+ Genotype of Experimental Strain: SOS-R1 NfsA ada(del) ogt(del)	Reference Condition: 5-nitrofuran nitrofurazone(5 micromolar for 3 hours compared to 0 micromolar)	OMP:0005023	presence of an SOS response	ECO:0000297	LacZ transcript localization evidence	Reference section 3.3; NfsA (Ec)= 2-NO2 nitroreductase only, from E. coli; phenotype is unchanged
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SOS-R1 83333	Genotype of Reference Strain: SOS-R1 NfsA ada+ ogt+ Genotype of Experimental Strain: SOS-R1 NfsA ada(del) ogt(del)	Reference Condition: 5-nitrofuran nitrofurazone(5 micromolar for 3 hours compared to 0 micromolar)	OMP:0005023	presence of an SOS response	ECO:0000297	LacZ transcript localization evidence	Reference section 3.3; NfsA (Kp)= 2-NO2 nitroreductase only, from K. pneumonia; phenotype is unchanged
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SOS-R1 83333	Genotype of Reference Strain: SOS-R1 NfsB ada+ ogt+ Genotype of Experimental Strain: SOS-R1 NfsB ada(del) ogt(del)	Reference Condition: 5-nitrofuran nitrofurazone(5 micromolar for 3 hours compared to 0 micromolar)	OMP:0005023	presence of an SOS response	ECO:0000297	LacZ transcript localization evidence	Reference section 3.3; NfsB= 2-NO2 or 4-NO2 nitroreductase; phenotype is unchanged
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SOS-R1 83333	Genotype of Reference Strain: SOS-R1 NemA ada+ ogt+ Genotype of Experimental Strain: SOS-R1 NemA ada(del) ogt(del)	Reference Condition: 5-nitrofuran nitrofurazone(5 micromolar for 3 hours compared to 0 micromolar)	OMP:0005023	presence of an SOS response	ECO:0000297	LacZ transcript localization evidence	Reference section 3.3; NemA= 4-NO2 nitroreductase only; phenotype is unchanged
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: SOS-R1 83333	Genotype of Reference Strain: SOS-R1 YfkO ada+ ogt+ Genotype of Experimental Strain: SOS-R1 YfkO ada(del) ogt(del)	Reference Condition: 5-nitrofuran nitrofurazone(5 micromolar for 3 hours compared to 0 micromolar)	OMP:0005023	presence of an SOS response	ECO:0000297	LacZ transcript localization evidence	Reference section 3.3; YfkO (Bs)= 4-NO2 nitroreductase only, from B. licheniformis; phenotype is unchanged
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References

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