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. 1993 Sep;175(17):5428–5437. doi: 10.1128/jb.175.17.5428-5437.1993

Metalloregulation in Bacillus subtilis: isolation and characterization of two genes differentially repressed by metal ions.

L Chen 1, L P James 1, J D Helmann 1
PMCID: PMC206598  PMID: 8396117

Abstract

We have cloned two metal-regulated genes (mrgA and mrgC) from Bacillus subtilis by using transposon Tn917-lacZ. Both were isolated as iron-repressible gene fusions, but the metal specificity and sensitivity of gene repression are distinct. Transcription of mrgA-lacZ is induced at the end of logarithmic-phase growth in minimal medium, and this induction is prevented by excess manganese, iron, cobalt, or copper. Limitation for metal ions is sufficient for mrgA-lacZ induction, since resuspension in medium lacking both manganese and iron rapidly induces transcription. Transcription of mrgC-lacZ is also induced by iron deprivation but is not repressed by added manganese or other metal ions. Expression of mrgC-lacZ and a 2,3-dihydroxybenzoic acid-based siderophore is repressed in parallel by iron, and in both cases, only iron effects repression. We have cloned and sequenced the promoter and regulatory regions of both mrgA and mrgC. Both genes are preceded by a predicted sigma A-dependent promoter element with overlapping sequences similar to the iron box consensus element for recognition by the Escherichia coli ferric uptake regulator protein (Fur). Mutation of the putative iron box for gene mrgC leads to partial derepression in iron-replete medium.

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Selected References

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  1. Bagg A., Neilands J. B. 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. 1987 Aug 25;26(17):5471–5477. doi: 10.1021/bi00391a039. [DOI] [PubMed] [Google Scholar]
  2. Bagg A., Neilands J. B. Molecular mechanism of regulation of siderophore-mediated iron assimilation. Microbiol Rev. 1987 Dec;51(4):509–518. doi: 10.1128/mr.51.4.509-518.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Blumentals I. I., Kelly R. M., Gorziglia M., Kaufman J. B., Shiloach J. Development of a defined medium and two-step culturing method for improved exotoxin A yields from Pseudomonas aeruginosa. Appl Environ Microbiol. 1987 Sep;53(9):2013–2020. doi: 10.1128/aem.53.9.2013-2020.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Boyd J., Oza M. N., Murphy J. R. Molecular cloning and DNA sequence analysis of a diphtheria tox iron-dependent regulatory element (dtxR) from Corynebacterium diphtheriae. Proc Natl Acad Sci U S A. 1990 Aug;87(15):5968–5972. doi: 10.1073/pnas.87.15.5968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Brickman T. J., Ozenberger B. A., McIntosh M. A. Regulation of divergent transcription from the iron-responsive fepB-entC promoter-operator regions in Escherichia coli. J Mol Biol. 1990 Apr 20;212(4):669–682. doi: 10.1016/0022-2836(90)90229-F. [DOI] [PubMed] [Google Scholar]
  6. Coy M., Neilands J. B. Structural dynamics and functional domains of the fur protein. Biochemistry. 1991 Aug 20;30(33):8201–8210. doi: 10.1021/bi00247a016. [DOI] [PubMed] [Google Scholar]
  7. Fisher S., Buxbaum L., Toth K., Eisenstadt E., Silver S. Regulation of manganese accumulation and exchange in Bacillus subtilis W23. J Bacteriol. 1973 Mar;113(3):1373–1380. doi: 10.1128/jb.113.3.1373-1380.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Griffiths E. Iron and bacterial virulence--a brief overview. Biol Met. 1991;4(1):7–13. doi: 10.1007/BF01135551. [DOI] [PubMed] [Google Scholar]
  9. Griggs D. W., Konisky J. Mechanism for iron-regulated transcription of the Escherichia coli cir gene: metal-dependent binding of fur protein to the promoters. J Bacteriol. 1989 Feb;171(2):1048–1054. doi: 10.1128/jb.171.2.1048-1054.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hantke K. Selection procedure for deregulated iron transport mutants (fur) in Escherichia coli K 12: fur not only affects iron metabolism. Mol Gen Genet. 1987 Nov;210(1):135–139. doi: 10.1007/BF00337769. [DOI] [PubMed] [Google Scholar]
  11. Hennecke H. Regulation of bacterial gene expression by metal-protein complexes. Mol Microbiol. 1990 Oct;4(10):1621–1628. doi: 10.1111/j.1365-2958.1990.tb00538.x. [DOI] [PubMed] [Google Scholar]
  12. Hoch J. A. Genetic analysis in Bacillus subtilis. Methods Enzymol. 1991;204:305–320. doi: 10.1016/0076-6879(91)04015-g. [DOI] [PubMed] [Google Scholar]
  13. Kunkel T. A., Roberts J. D., Zakour R. A. Rapid and efficient site-specific mutagenesis without phenotypic selection. Methods Enzymol. 1987;154:367–382. doi: 10.1016/0076-6879(87)54085-x. [DOI] [PubMed] [Google Scholar]
  14. Litwin C. M., Boyko S. A., Calderwood S. B. Cloning, sequencing, and transcriptional regulation of the Vibrio cholerae fur gene. J Bacteriol. 1992 Mar;174(6):1897–1903. doi: 10.1128/jb.174.6.1897-1903.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Love P. E., Lyle M. J., Yasbin R. E. DNA-damage-inducible (din) loci are transcriptionally activated in competent Bacillus subtilis. Proc Natl Acad Sci U S A. 1985 Sep;82(18):6201–6205. doi: 10.1073/pnas.82.18.6201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Martínez J. L., Delgado-Iribarren A., Baquero F. Mechanisms of iron acquisition and bacterial virulence. FEMS Microbiol Rev. 1990 Mar;6(1):45–56. doi: 10.1111/j.1574-6968.1990.tb04085.x. [DOI] [PubMed] [Google Scholar]
  17. Moran C. P., Jr, Lang N., LeGrice S. F., Lee G., Stephens M., Sonenshein A. L., Pero J., Losick R. Nucleotide sequences that signal the initiation of transcription and translation in Bacillus subtilis. Mol Gen Genet. 1982;186(3):339–346. doi: 10.1007/BF00729452. [DOI] [PubMed] [Google Scholar]
  18. Neidhardt F. C., Bloch P. L., Smith D. F. Culture medium for enterobacteria. J Bacteriol. 1974 Sep;119(3):736–747. doi: 10.1128/jb.119.3.736-747.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Ozcengiz G., Alaeddinoglu N. G., Demain A. L. Regulation of biosynthesis of bacilysin by Bacillus subtilis. J Ind Microbiol. 1990 Oct;6(2):91–100. doi: 10.1007/BF01576428. [DOI] [PubMed] [Google Scholar]
  20. Payne S. M. Iron and virulence in Shigella. Mol Microbiol. 1989 Sep;3(9):1301–1306. doi: 10.1111/j.1365-2958.1989.tb00281.x. [DOI] [PubMed] [Google Scholar]
  21. Peters W. J., Warren R. A. Itoic acid synthesis in Bacillus subtilis. J Bacteriol. 1968 Feb;95(2):360–366. doi: 10.1128/jb.95.2.360-366.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Peters W. J., Warren R. A. Phenolic acids and iron transport in Bacillus subtilis. Biochim Biophys Acta. 1968 Sep 3;165(2):225–232. doi: 10.1016/0304-4165(68)90050-0. [DOI] [PubMed] [Google Scholar]
  23. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Schmitt M. P., Holmes R. K. Iron-dependent regulation of diphtheria toxin and siderophore expression by the cloned Corynebacterium diphtheriae repressor gene dtxR in C. diphtheriae C7 strains. Infect Immun. 1991 Jun;59(6):1899–1904. doi: 10.1128/iai.59.6.1899-1904.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Schmitt M. P., Twiddy E. M., Holmes R. K. Purification and characterization of the diphtheria toxin repressor. Proc Natl Acad Sci U S A. 1992 Aug 15;89(16):7576–7580. doi: 10.1073/pnas.89.16.7576. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Schneider R., Hantke K. Iron-hydroxamate uptake systems in Bacillus subtilis: identification of a lipoprotein as part of a binding protein-dependent transport system. Mol Microbiol. 1993 Apr;8(1):111–121. doi: 10.1111/j.1365-2958.1993.tb01208.x. [DOI] [PubMed] [Google Scholar]
  27. Silver S., Walderhaug M. Gene regulation of plasmid- and chromosome-determined inorganic ion transport in bacteria. Microbiol Rev. 1992 Mar;56(1):195–228. doi: 10.1128/mr.56.1.195-228.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Staggs T. M., Perry R. D. Fur regulation in Yersinia species. Mol Microbiol. 1992 Sep;6(17):2507–2516. doi: 10.1111/j.1365-2958.1992.tb01427.x. [DOI] [PubMed] [Google Scholar]
  29. Summers A. O. Untwist and shout: a heavy metal-responsive transcriptional regulator. J Bacteriol. 1992 May;174(10):3097–3101. doi: 10.1128/jb.174.10.3097-3101.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Tao X., Boyd J., Murphy J. R. Specific binding of the diphtheria tox regulatory element DtxR to the tox operator requires divalent heavy metal ions and a 9-base-pair interrupted palindromic sequence. Proc Natl Acad Sci U S A. 1992 Jul 1;89(13):5897–5901. doi: 10.1073/pnas.89.13.5897. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Van Hove B., Staudenmaier H., Braun V. Novel two-component transmembrane transcription control: regulation of iron dicitrate transport in Escherichia coli K-12. J Bacteriol. 1990 Dec;172(12):6749–6758. doi: 10.1128/jb.172.12.6749-6758.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Vander Horn P. B., Zahler S. A. Cloning and nucleotide sequence of the leucyl-tRNA synthetase gene of Bacillus subtilis. J Bacteriol. 1992 Jun;174(12):3928–3935. doi: 10.1128/jb.174.12.3928-3935.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Vellanoweth R. L., Rabinowitz J. C. The influence of ribosome-binding-site elements on translational efficiency in Bacillus subtilis and Escherichia coli in vivo. Mol Microbiol. 1992 May;6(9):1105–1114. doi: 10.1111/j.1365-2958.1992.tb01548.x. [DOI] [PubMed] [Google Scholar]
  34. Walsh B. L., Peters W. J., Warren R. A. The regulation of phenolic acid sysdtness in Bacillus subtilis. Can J Microbiol. 1971 Jan;17(1):53–59. doi: 10.1139/m71-009. [DOI] [PubMed] [Google Scholar]
  35. Weinberg E. D. Roles of trace metals in transcriptional control of microbial secondary metabolism. Biol Met. 1990;2(4):191–196. doi: 10.1007/BF01141358. [DOI] [PubMed] [Google Scholar]
  36. von Hippel P. H., Berg O. G. On the specificity of DNA-protein interactions. Proc Natl Acad Sci U S A. 1986 Mar;83(6):1608–1612. doi: 10.1073/pnas.83.6.1608. [DOI] [PMC free article] [PubMed] [Google Scholar]

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