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J Bacteriol. Aug 1990; 172(8): 4197–4205.
PMCID: PMC213242

soxR, a locus governing a superoxide response regulon in Escherichia coli K-12.

Abstract

The nfo (endonuclease IV) gene of Escherichia coli is induced by superoxide generators such as paraquat (methyl viologen). An nfo'-lacZ operon fusion was used to isolate extragenic mutations affecting its expression. The mutations also affected the expression of glucose 6-phosphate dehydrogenase, Mn2(+)-superoxide dismutase (sodA), and three lacZ fusions to soi (superoxide-inducible) genes of unknown function. The mutations were located 2 kilobases clockwise of ssb at 92 min on the current linkage map. One set of mutations, in a new gene designated soxR, caused constitutive overexpression of nfo and the other genes. It included insertions or deletions affecting the carboxyl end of a 17-kilodalton polypeptide. In a soxR mutant, the expression of sodA, unlike that of nfo, was also regulated independently by oxygen tension. Two other mutants were isolated in which the target genes were noninducible; they had an increased sensitivity to killing by superoxide-generating compounds. One had a Tn10 insertion in or near soxR; the other had a multigene deletion encompassing soxR. Therefore, the region functions as a positive regulator because it encodes one or more products needed for the induction of nfo. Regulation is likely to be at the level of transcription because the mutations were able to affect the expression of an nfo'-lac operon fusion that contained the ribosome-binding site for lacZ. Some mutant plasmids that failed to suppress (or complement) constitutivity in trans had insertion mutations several hundred nucleotides upstream of soxR in the general region of a gene for a 13-kilodalton protein encoded by the opposite strand, raising the possibility of a second regulatory gene in this region. The result define a new regulon, controlled by soxR, mediating at least part of the global response to superoxide in E. coli.

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  • Alton NK, Vapnek D. Nucleotide sequence analysis of the chloramphenicol resistance transposon Tn9. Nature. 1979 Dec 20;282(5741):864–869. [PubMed]
  • Anderson RP, Roth JR. Tandem genetic duplications in phage and bacteria. Annu Rev Microbiol. 1977;31:473–505. [PubMed]
  • Bachmann BJ. Pedigrees of some mutant strains of Escherichia coli K-12. Bacteriol Rev. 1972 Dec;36(4):525–557. [PMC free article] [PubMed]
  • Bachmann BJ. Linkage map of Escherichia coli K-12, edition 7. Microbiol Rev. 1983 Jun;47(2):180–230. [PMC free article] [PubMed]
  • Barany F. Two-codon insertion mutagenesis of plasmid genes by using single-stranded hexameric oligonucleotides. Proc Natl Acad Sci U S A. 1985 Jun;82(12):4202–4206. [PMC free article] [PubMed]
  • Bittner M, Vapnek D. Versatile cloning vectors derived from the runaway-replication plasmid pKN402. Gene. 1981 Dec;15(4):319–329. [PubMed]
  • Brawn MK, Fridovich I. Increased superoxide radical production evokes inducible DNA repair in Escherichia coli. J Biol Chem. 1985 Jan 25;260(2):922–925. [PubMed]
  • Carlioz A, Touati D. Isolation of superoxide dismutase mutants in Escherichia coli: is superoxide dismutase necessary for aerobic life? EMBO J. 1986 Mar;5(3):623–630. [PMC free article] [PubMed]
  • Chan E, Weiss B. Endonuclease IV of Escherichia coli is induced by paraquat. Proc Natl Acad Sci U S A. 1987 May;84(10):3189–3193. [PMC free article] [PubMed]
  • Clarke L, Carbon J. A colony bank containing synthetic Col El hybrid plasmids representative of the entire E. coli genome. Cell. 1976 Sep;9(1):91–99. [PubMed]
  • Demple B, Johnson A, Fung D. Exonuclease III and endonuclease IV remove 3' blocks from DNA synthesis primers in H2O2-damaged Escherichia coli. Proc Natl Acad Sci U S A. 1986 Oct;83(20):7731–7735. [PMC free article] [PubMed]
  • Fridovich I. Superoxide radical: an endogenous toxicant. Annu Rev Pharmacol Toxicol. 1983;23:239–257. [PubMed]
  • Greenberg JT, Demple B. A global response induced in Escherichia coli by redox-cycling agents overlaps with that induced by peroxide stress. J Bacteriol. 1989 Jul;171(7):3933–3939. [PMC free article] [PubMed]
  • Guyer MS. The gamma delta sequence of F is an insertion sequence. J Mol Biol. 1978 Dec 15;126(3):347–365. [PubMed]
  • Guyer MS. Uses of the transposon gamma delta in the analysis of cloned genes. Methods Enzymol. 1983;101:362–369. [PubMed]
  • Hassan HM, Fridovich I. Enzymatic defenses against the toxicity of oxygen and of streptonigrin in Escherichia coli. J Bacteriol. 1977 Mar;129(3):1574–1583. [PMC free article] [PubMed]
  • Hassan HM, Fridovich I. Regulation of the synthesis of superoxide dismutase in Escherichia coli. Induction by methyl viologen. J Biol Chem. 1977 Nov 10;252(21):7667–7672. [PubMed]
  • Hassan HM, Fridovich I. Intracellular production of superoxide radical and of hydrogen peroxide by redox active compounds. Arch Biochem Biophys. 1979 Sep;196(2):385–395. [PubMed]
  • Kao SM, Hassan HM. Biochemical characterization of a paraquat-tolerant mutant of Escherichia coli. J Biol Chem. 1985 Sep 5;260(19):10478–10481. [PubMed]
  • Kogoma T, Farr SB, Joyce KM, Natvig DO. Isolation of gene fusions (soi::lacZ) inducible by oxidative stress in Escherichia coli. Proc Natl Acad Sci U S A. 1988 Jul;85(13):4799–4803. [PMC free article] [PubMed]
  • Kohara Y, Akiyama K, Isono K. The physical map of the whole E. coli chromosome: application of a new strategy for rapid analysis and sorting of a large genomic library. Cell. 1987 Jul 31;50(3):495–508. [PubMed]
  • Kushner SR, Nagaishi H, Clark AJ. Indirect suppression of recB and recC mutations by exonuclease I deficiency. Proc Natl Acad Sci U S A. 1972 Jun;69(6):1366–1370. [PMC free article] [PubMed]
  • Levin JD, Johnson AW, Demple B. Homogeneous Escherichia coli endonuclease IV. Characterization of an enzyme that recognizes oxidative damage in DNA. J Biol Chem. 1988 Jun 15;263(17):8066–8071. [PubMed]
  • Ljungquist S. A new endonuclease from Escherichia coli acting at apurinic sites in DNA. J Biol Chem. 1977 May 10;252(9):2808–2814. [PubMed]
  • Lopilato J, Bortner S, Beckwith J. Mutations in a new chromosomal gene of Escherichia coli K-12, pcnB, reduce plasmid copy number of pBR322 and its derivatives. Mol Gen Genet. 1986 Nov;205(2):285–290. [PubMed]
  • Mead DA, Szczesna-Skorupa E, Kemper B. Single-stranded DNA 'blue' T7 promoter plasmids: a versatile tandem promoter system for cloning and protein engineering. Protein Eng. 1986 Oct-Nov;1(1):67–74. [PubMed]
  • Morgan RW, Christman MF, Jacobson FS, Storz G, Ames BN. Hydrogen peroxide-inducible proteins in Salmonella typhimurium overlap with heat shock and other stress proteins. Proc Natl Acad Sci U S A. 1986 Nov;83(21):8059–8063. [PMC free article] [PubMed]
  • Ohtsubo H, Ohtsubo E. Nucleotide sequence of an insertion element, IS1. Proc Natl Acad Sci U S A. 1978 Feb;75(2):615–619. [PMC free article] [PubMed]
  • Sancar A, Hack AM, Rupp WD. Simple method for identification of plasmid-coded proteins. J Bacteriol. 1979 Jan;137(1):692–693. [PMC free article] [PubMed]
  • Sancar A, Wharton RP, Seltzer S, Kacinski BM, Clarke ND, Rupp WD. Identification of the uvrA gene product. J Mol Biol. 1981 May 5;148(1):45–62. [PubMed]
  • Sancar A, Williams KR, Chase JW, Rupp WD. Sequences of the ssb gene and protein. Proc Natl Acad Sci U S A. 1981 Jul;78(7):4274–4278. [PMC free article] [PubMed]
  • Saporito SM, Cunningham RP. Nucleotide sequence of the nfo gene of Escherichia coli K-12. J Bacteriol. 1988 Nov;170(11):5141–5145. [PMC free article] [PubMed]
  • Shapira SK, Chou J, Richaud FV, Casadaban MJ. New versatile plasmid vectors for expression of hybrid proteins coded by a cloned gene fused to lacZ gene sequences encoding an enzymatically active carboxy-terminal portion of beta-galactosidase. Gene. 1983 Nov;25(1):71–82. [PubMed]
  • Shimada K, Weisberg RA, Gottesman ME. Prophage lambda at unusual chromosomal locations. I. Location of the secondary attachment sites and the properties of the lysogens. J Mol Biol. 1972 Feb 14;63(3):483–503. [PubMed]
  • Simons RW, Houman F, Kleckner N. Improved single and multicopy lac-based cloning vectors for protein and operon fusions. Gene. 1987;53(1):85–96. [PubMed]
  • Singer M, Baker TA, Schnitzler G, Deischel SM, Goel M, Dove W, Jaacks KJ, Grossman AD, Erickson JW, Gross CA. A collection of strains containing genetically linked alternating antibiotic resistance elements for genetic mapping of Escherichia coli. Microbiol Rev. 1989 Mar;53(1):1–24. [PMC free article] [PubMed]
  • Smith PK, Krohn RI, Hermanson GT, Mallia AK, Gartner FH, Provenzano MD, Fujimoto EK, Goeke NM, Olson BJ, Klenk DC. Measurement of protein using bicinchoninic acid. Anal Biochem. 1985 Oct;150(1):76–85. [PubMed]
  • Smith TF, Sadler JR. The nature of lactose operator constitive mutations. J Mol Biol. 1971 Jul 28;59(2):273–305. [PubMed]
  • Studier FW, Moffatt BA. Use of bacteriophage T7 RNA polymerase to direct selective high-level expression of cloned genes. J Mol Biol. 1986 May 5;189(1):113–130. [PubMed]
  • Sutcliffe JG. Complete nucleotide sequence of the Escherichia coli plasmid pBR322. Cold Spring Harb Symp Quant Biol. 1979;43(Pt 1):77–90. [PubMed]
  • Tabor S, Richardson CC. A bacteriophage T7 RNA polymerase/promoter system for controlled exclusive expression of specific genes. Proc Natl Acad Sci U S A. 1985 Feb;82(4):1074–1078. [PMC free article] [PubMed]
  • Taylor AF, Siliciano PG, Weiss B. Cloning of the dut (deoxyuridine triphosphatase) gene of Escherichia coli. Gene. 1980 May;9(3-4):321–336. [PubMed]
  • Walkup LK, Kogoma T. Escherichia coli proteins inducible by oxidative stress mediated by the superoxide radical. J Bacteriol. 1989 Mar;171(3):1476–1484. [PMC free article] [PubMed]
  • Way JC, Davis MA, Morisato D, Roberts DE, Kleckner N. New Tn10 derivatives for transposon mutagenesis and for construction of lacZ operon fusions by transposition. Gene. 1984 Dec;32(3):369–379. [PubMed]
  • White BJ, Hochhauser SJ, Cintron NM, Weiss B. Genetic mapping of xthA, the structural gene for exonuclease III in Escherichia coli K-12. J Bacteriol. 1976 Jun;126(3):1082–1088. [PMC free article] [PubMed]
  • Winans SC, Elledge SJ, Krueger JH, Walker GC. Site-directed insertion and deletion mutagenesis with cloned fragments in Escherichia coli. J Bacteriol. 1985 Mar;161(3):1219–1221. [PMC free article] [PubMed]

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