1887

Microbiology Society logo

Volume 139, Issue 11

Characterization of regulatory mutations causing anaerobic derepression of the gene in K12: cooperation between - and -acting regulatory loci Free

Abstract

The genetic loci leading to anaerobic derepression of a protein fusion in a UV-generated mutant strain (UV14) of were identified. The mutant (UV14) was found to harbour two altered loci: one is in the -regulatory gene (fumarate nitrate reduction) where leucine-129 was changed to glutamine (), and the second () is in the promoter region () of the gene apparently affecting the binding of the Fur (ferric uptake regulation) protein. Introduction of an gene into UV14 restored anaerobic repression of and restored the ability of the cells to reduce nitrate. However, when either the or the mutation was introduced into an otherwise wild-type background, only slight anaerobic derepression of was observed. When both the - and -acting mutations (i.e. and ) were combined simultaneously in an otherwise wild-type background, the specific activity of expression was comparable to that of the original mutant strain (UV14). Furthermore, a genetically confirmed double mutant was also similarly derepressed in anaerobic expression. The data presented suggest that the -mutation in UV14 () affects the Fur-binding site in the promoter, while having no effect on Fnr or Arc mediated repression. Also, a second putative Fnr-binding site that straddles the ribosomal binding-site was identified in the gene.

  • Received:
  • Accepted:
  • Revised:
  • Published Online:
© Society for General Microbiology 1993
Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-139-11-2677
1993-11-01
2024-04-20
Loading full text...

Full text loading...

/deliver/fulltext/micro/139/11/mic-139-11-2677.html?itemId=/content/journal/micro/10.1099/00221287-139-11-2677&mimeType=html&fmt=ahah

References

  1. Bachmann B. J. 1990; Linkage map of Escherichia coli K12, edition 8. Microbiological Reviews 54:130–197
     [Google Scholar]
  2. Bagg A., Neilands J. B. 1987; Ferric uptake regulation protein acts as a repressor, employing iron(II) as a cofactor to bind the operator of an iron transport operon in Escherichia coli. Biochemistry 26:5471–5477
     [Google Scholar]
  3. Beauchamp C. O., Fridovich I. 1971; Superoxide dismutase: improved assays and an assay applicable to acrylamide gels. Analytical Biochemistry 44:276–287
     [Google Scholar]
  4. Beaumont M., Hassan H. M. 1990; Role of fur regulon on the expression of sodA in Escherichia coli. Abstr. H-114 p. 173: Abstr. 90th Annual Meeting, American Society for Microbiology.
    [Google Scholar]
  5. Beaumont M., Hassan H. M. 1991; Characterization of a sodA mutant of Escherichia coli: regulatory role of Arc and Fur. FASEB Journal (Abstract) 5:2586
     [Google Scholar]
  6. Beaumont M., Hassan H. M. 1992; Characterization of a sodA mutant of Escherichia coli. Current Microbiology 25:135–141
     [Google Scholar]
  7. Compan I., Touati D. 1993; Interaction of six global transcription regulators in expression of manganese superoxide dismutase in Escherichia coli K12. Journal of Bacteriology 175:1687–1696
     [Google Scholar]
  8. Gardner P., Fridovich I. 1987; Controls on the biosynthesis of the manganese-containing superoxide dismutase of Escherichia coli. Journal of Biological Chemistry 262:17591–17595
     [Google Scholar]
  9. Greenberg J. T., Monach P. A., Chou J. -H., Josephy P. D., Demple B. 1990; Positive control of a global antioxidant defense regulon activated by superoxide-generating agents in Escherichia coli. Proceedings of the National Academy of Sciences of the United States of America 87:6181–6185
     [Google Scholar]
  10. Gregory E. M., Fridovich I. 1973; The induction of superoxide dismutase by molecular oxygen. Journal of Bacteriology 114:543–548
     [Google Scholar]
  11. Hanson R. S., Phillips J. A. 1981; Chemical composition. In Manual of Methods for General Bacteriology p. 355 Washington, DC: American Society for Microbiology;
     [Google Scholar]
  12. Hassan H. M. 1989; Microbial superoxide dismutases. Advances in Genetics 26:65–97
     [Google Scholar]
  13. Hassan H. M., Fridovich I. 1977; Regulation of the synthesis of superoxide dismutase in Escherichia coli: induction by methyl viologen. Journal of Biological Chemistry 252:7667–7672
     [Google Scholar]
  14. Hassan H. M., Fridovich I. 1979; Intracellular production of superoxide radical and of hydrogen peroxide by redox active compounds. Archives of Biochemistry and Biophysics 196:385–395
     [Google Scholar]
  15. Hassan H. M., Moody C. S. 1984; Induction of the manganese-containing superoxide dismutase in Escherichia coli by nalidixic acid and iron chelators. FEMS Microbioloty Letters 25:233–236
     [Google Scholar]
  16. Hassan H. M., Sun H. C. H. 1991; Regulatory role of Fnr in the expression of Mn-superoxide dismutase in Escherichia coli. FASEB Journal (Abstract) 5:2585
     [Google Scholar]
  17. Hassan H. M., Sun H. C. H. 1992; Regulatory roles of Fnr, Fur and Arc in the expression of the manganese containing superoxide dismutase in Escherichia coli. Proceedings of the National Academy of Sciences of the United States of America 89:3217–3221
     [Google Scholar]
  18. Iuchi S., Cameron D., Lin E. C. C. 1989; A second global regulator gene (arcB) mediates repression of enzymes in aerobic pathways of Escherichia coli. Journal of Bacteriology 171:868–873
     [Google Scholar]
  19. Jones H. M., Gunsalus R. P. 1987; Regulation of Escherichia colifumarate reductase operon expression by resP1ratory electron acceptors and the fnr gene product. Journal of Bacteriology 169:3340–3349
     [Google Scholar]
  20. Kiley P. J., Reznikoff W. S. 1991; Fnr mutants that activate gene expression in the presence of oxygen. Journal of Bacteriology 173:16–22
     [Google Scholar]
  21. Mccord J., Fridovich I. 1969; Superoxide dismutase: an enzymatic function for erythrocuprein (hemocuprein). Journal of Biological Chemistry 244:6049–6055
     [Google Scholar]
  22. Miller J. 1972 Experiments in Molecular Genetics Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
     [Google Scholar]
  23. Moody C. S., Hassan H. M. 1984; Anaerobic biosynthesis of the manganese-containing superoxide dismutase in Escherichia coli. Journal of Biological Chemistry 259:12821–12825
     [Google Scholar]
  24. Naik S., Hassan H. M. 1990; Use of site directed mutagenesis to identify an upstream regulatory sequence of sodA gene of Escherichia coli K12. Proceedings of the National Academy of Sciences of the United States of America 87:2618–2622
     [Google Scholar]
  25. Neiderhoffer E., Naranjo C., Fee J. 1989; Relationship of the superoxide dismutase genes (sodA and sodB) to the iron uptake (fur) in Escherichia coli K12. UCLA Symposium on Molecular and Cellular Biology 98:149–158
     [Google Scholar]
  26. Neiderhoffer E., Naranjo C., Bradley K., Fee J. 1990; Control of Escherichia coli superoxide dismutase (sodA and sodB) genes by the ferric uptake regulation (fur) locus. Journal of Bacteriology 172:1930–1938
     [Google Scholar]
  27. Presutti D. G. 1992; Identification of sodA sequence-specific binding proteins in Escherichia coli. abstr. H-310 p. 234: Abstr. 92nd Annual Meeting, American Society for Microbiology.
    [Google Scholar]
  28. Privalle C., Beyer W. F., Fridovich I. 1989; Anaerobic induction of ProMn-superoxide dismutase in Escherichia coli. Journal of Biological Chemistry 264:2758–2763
     [Google Scholar]
  29. Pugh S., Fridovich I. 1985; Induction of superoxide dismutases in Escherichia coli B by metal chelators. Journal of Bacteriology 162:196–202
     [Google Scholar]
  30. Sambrook J., Fritsch E., Maniatis T. 1989 Molecular Cloning: A Laboratory Manual, 2nd edn.. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
     [Google Scholar]
  31. Sanger F., Nicklen S., Coulsen A. R. 1977; DNA sequencing with chain terminating inhibitors. Proceedings of the National Academy of Sciences of the United States of America 74:5463–5467
     [Google Scholar]
  32. Sawers G., Suppmann B. 1992; Anaerobic induction of pyruvate formate-lyase gene expression is mediated by the ArcA and Fnr proteins. Journal of Bacteriology 174:3474–3478
     [Google Scholar]
  33. Schiavone J. R., Hassan H. M. 1988; The role of redox in the regulation of manganese-containing superoxide dismutase biosynthesis in Escherichia coli. Journal of Biological Chemistry 263:4269–4273
     [Google Scholar]
  34. Schrum L. W., Hassan H. M. 1990; Role of DNA topology in the expression of the sodA gene in Escherichia coli using operon fusion. Abstract H-73 p. 166 Abstr. 90th Annual Meeting, American Society for Microbiology.
    [Google Scholar]
  35. Schrum L. W., Hassan H. M. 1992; Transcriptional regulation of Mn-superoxide dismutase gene (sodA) of Escherichia coli is stimulated by DNA gyrase inhibitors. Archives of Biochemistry and Biophysics 299:185–192
     [Google Scholar]
  36. Singer M., Baker T., Schnitzler G., Deischel S., Goel M., Dove W., Jaacks K., Grossman A., Erickson J., Gross C. 1989; A collection of strains containing genetically linked alternating antibiotic resistance elements for genetic mapP1ng of Escherichia coli. Microbiological Reviews 53:1–24
     [Google Scholar]
  37. Shaw S., Guest J. R. 1982; Nucleotide sequence of the fnr gene and primary structure of the Fnr protein of Escherichia coli. Nucleic Acids Research 10:6119–6130
     [Google Scholar]
  38. Spiro S., Guest J. R. 1990; Fnr and its role in oxygen regulated gene expression in Escherichia coli. FEMS Microbiology Reviews 75:399–428
     [Google Scholar]
  39. Takeda Y., Avila H. 1986; Structure and gene expression of the Escherichia coli manganese superoxide dismutase gene. Nucleic Acids Research 14:4577–4589
     [Google Scholar]
  40. Tardat B., Tuoati D. 1991; Two global regulators repress the anaerobic expression of MnSOD in E. coli: Fur (ferric uptake regulation) and Arc (aerobic resP1ratory control). Molecular Microbiology 5:455–466
     [Google Scholar]
  41. Touati D. 1983; Cloning and mapP1ng of the manganese superoxide gene (sodA) of Escherichia coli K12. Journal of Bacteriology 155:1078–1087
     [Google Scholar]
  42. Touati D. 1988; Transcriptional and post-transcriptional regulation of manganese superoxide dismutase biosynthesis in Escherichia coli studied with operon and protein fusions. Journal of Bacteriology 170:2511–2520
     [Google Scholar]
  43. Trageser M., Spiro S., Duchene A., Kojro E., Fahrenholz F., Guest J. R., Unden G. 1990; Isolation of intact Fnr protein of Escherichia coli. Molecular Microbiology 4:21–27
     [Google Scholar]
  44. Tsaneva I., Weiss B. 1990; soxR, a locus governing a superoxide response in Escherichia coli K12. Journal of Bacteriology 172:4197–4205
     [Google Scholar]
  45. Unden G., Trageser M. 1991; Oxygen regulated gene expression in Escherichia coli: control of anaerobic resP1ration by the Fnr protein. Antonie van Leeuwenhoek 59:65–76
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-139-11-2677
Loading
Characterization of regulatory mutations causing anaerobic derepression of the sodA gene in Escherichia coli K12: cooperation between cis- and trans-acting regulatory loci
Microbiology 139, 2677 (1993); https://doi.org/10.1099/00221287-139-11-2677
/content/journal/micro/10.1099/00221287-139-11-2677
/content/journal/micro/10.1099/00221287-139-11-2677
Loading

Data & Media loading...

Most cited Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error