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. 1996 Feb;178(3):854–861. doi: 10.1128/jb.178.3.854-861.1996

Duplicate isochorismate synthase genes of Bacillus subtilis: regulation and involvement in the biosyntheses of menaquinone and 2,3-dihydroxybenzoate.

B M Rowland 1, H W Taber 1
PMCID: PMC177735  PMID: 8550523

Abstract

Bacillus subtilis has duplicate isochorismate synthase genes, menF and dhbC. Isochorismate synthase is involved in the biosynthesis of both the respiratory chain component menaquinone (MK) and the siderophore 2,3-dihydroxybenzoate (DHB). Several menF and dhbC deletion mutants were constructed to identify the contribution made by each gene product to MK and DHB biosynthesis. menF deletion mutants were able to produce wild-type levels of MK and DHB, suggesting that the dhbC gene product is able to compensate for the lack of MenF. However, a dhbC deletion mutant produced wild-type levels of MK but was DHB deficient, indicating that MenF is unable to compensate for the lack of DhbC. A menF dhbC double-deletion mutant was both MK and DHB deficient. Transcription analysis showed that expression of dhbC, but not of menF, is regulated by iron concentration. A dhbA'::lacZ fusion strain was constructed to examine the effects of mutations to the iron box sequence within the dhb promoter region. These mutations abolished the iron-regulated transcription of the dhb genes, suggesting that a Fur-like repressor protein exists in B. subtilis.

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Selected References

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  1. 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]
  2. Chen L., James L. P., Helmann J. D. Metalloregulation in Bacillus subtilis: isolation and characterization of two genes differentially repressed by metal ions. J Bacteriol. 1993 Sep;175(17):5428–5437. doi: 10.1128/jb.175.17.5428-5437.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Dahl M. K., Meinhof C. G. A series of integrative plasmids for Bacillus subtilis containing unique cloning sites in all three open reading frames for translational lacZ fusions. Gene. 1994 Jul 22;145(1):151–152. doi: 10.1016/0378-1119(94)90341-7. [DOI] [PubMed] [Google Scholar]
  4. Grossman T. H., Tuckman M., Ellestad S., Osburne M. S. Isolation and characterization of Bacillus subtilis genes involved in siderophore biosynthesis: relationship between B. subtilis sfpo and Escherichia coli entD genes. J Bacteriol. 1993 Oct;175(19):6203–6211. doi: 10.1128/jb.175.19.6203-6211.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Günter K., Toupet C., Schupp T. Characterization of an iron-regulated promoter involved in desferrioxamine B synthesis in Streptomyces pilosus: repressor-binding site and homology to the diphtheria toxin gene promoter. J Bacteriol. 1993 Jun;175(11):3295–3302. doi: 10.1128/jb.175.11.3295-3302.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hill K. F., Mueller J. P., Taber H. W. The Bacillus subtilis menCD promoter is responsive to extracellular pH. Arch Microbiol. 1990;153(4):355–359. doi: 10.1007/BF00249005. [DOI] [PubMed] [Google Scholar]
  7. Ho S. N., Hunt H. D., Horton R. M., Pullen J. K., Pease L. R. Site-directed mutagenesis by overlap extension using the polymerase chain reaction. Gene. 1989 Apr 15;77(1):51–59. doi: 10.1016/0378-1119(89)90358-2. [DOI] [PubMed] [Google Scholar]
  8. Hulett F. M., Bookstein C., Jensen K. Evidence for two structural genes for alkaline phosphatase in Bacillus subtilis. J Bacteriol. 1990 Feb;172(2):735–740. doi: 10.1128/jb.172.2.735-740.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hulett F. M., Kim E. E., Bookstein C., Kapp N. V., Edwards C. W., Wyckoff H. W. Bacillus subtilis alkaline phosphatases III and IV. Cloning, sequencing, and comparisons of deduced amino acid sequence with Escherichia coli alkaline phosphatase three-dimensional structure. J Biol Chem. 1991 Jan 15;266(2):1077–1084. [PubMed] [Google Scholar]
  10. Jin S., Sonenshein A. L. Identification of two distinct Bacillus subtilis citrate synthase genes. J Bacteriol. 1994 Aug;176(15):4669–4679. doi: 10.1128/jb.176.15.4669-4679.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Jin S., Sonenshein A. L. Transcriptional regulation of Bacillus subtilis citrate synthase genes. J Bacteriol. 1994 Aug;176(15):4680–4690. doi: 10.1128/jb.176.15.4680-4690.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Martinussen J., Glaser P., Andersen P. S., Saxild H. H. Two genes encoding uracil phosphoribosyltransferase are present in Bacillus subtilis. J Bacteriol. 1995 Jan;177(1):271–274. doi: 10.1128/jb.177.1.271-274.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Meganathan R., Coffell R. Identity of the quinone in Bacillus alcalophilus. J Bacteriol. 1985 Nov;164(2):911–913. doi: 10.1128/jb.164.2.911-913.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Miller P., Mueller J., Hill K., Taber H. Transcriptional regulation of a promoter in the men gene cluster of Bacillus subtilis. J Bacteriol. 1988 Jun;170(6):2742–2748. doi: 10.1128/jb.170.6.2742-2748.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Miller P., Rabinowitz A., Taber H. Molecular cloning and preliminary genetic analysis of the men gene cluster of Bacillus subtilis. J Bacteriol. 1988 Jun;170(6):2735–2741. doi: 10.1128/jb.170.6.2735-2741.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Neuhard J., Price A. R., Schack L., Thomassen E. Two thymidylate synthetases in Bacillus subtilis. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1194–1198. doi: 10.1073/pnas.75.3.1194. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Oguiza J. A., Tao X., Marcos A. T., Martín J. F., Murphy J. R. Molecular cloning, DNA sequence analysis, and characterization of the Corynebacterium diphtheriae dtxR homolog from Brevibacterium lactofermentum. J Bacteriol. 1995 Jan;177(2):465–467. doi: 10.1128/jb.177.2.465-467.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. 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]
  19. 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]
  20. Piggot P. J., Curtis C. A., de Lencastre H. Use of integrational plasmid vectors to demonstrate the polycistronic nature of a transcriptional unit (spoIIA) required for sporulation of Bacillus subtilis. J Gen Microbiol. 1984 Aug;130(8):2123–2136. doi: 10.1099/00221287-130-8-2123. [DOI] [PubMed] [Google Scholar]
  21. Qin X., Taber H. W. Transcriptional regulation of the Bacillus subtilis menp1 promoter. J Bacteriol. 1996 Feb;178(3):705–713. doi: 10.1128/jb.178.3.705-713.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Salton M. R., Schmitt M. D. Effects of diphenylamine on carotenoids and menaquinones in bacterial membranes. Biochim Biophys Acta. 1967 May 2;135(2):196–207. doi: 10.1016/0005-2736(67)90114-9. [DOI] [PubMed] [Google Scholar]
  23. Saraste M., Metso T., Nakari T., Jalli T., Lauraeus M., Van der Oost J. The Bacillus subtilis cytochrome-c oxidase. Variations on a conserved protein theme. Eur J Biochem. 1991 Jan 30;195(2):517–525. doi: 10.1111/j.1432-1033.1991.tb15732.x. [DOI] [PubMed] [Google Scholar]
  24. Schaeffer P., Millet J., Aubert J. P. Catabolic repression of bacterial sporulation. Proc Natl Acad Sci U S A. 1965 Sep;54(3):704–711. doi: 10.1073/pnas.54.3.704. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. 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]
  26. Stojiljkovic I., Bäumler A. J., Hantke K. Fur regulon in gram-negative bacteria. Identification and characterization of new iron-regulated Escherichia coli genes by a fur titration assay. J Mol Biol. 1994 Feb 18;236(2):531–545. doi: 10.1006/jmbi.1994.1163. [DOI] [PubMed] [Google Scholar]
  27. Sugahara T., Konno Y., Ohta H., Ito K., Kaneko J., Kamio Y., Izaki K. Purification and properties of two membrane alkaline phosphatases from Bacillus subtilis 168. J Bacteriol. 1991 Mar;173(5):1824–1826. doi: 10.1128/jb.173.5.1824-1826.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Taber H. W., Dellers E. A., Lombardo L. R. Menaquinone biosynthesis in Bacillus subtilis: isolation of men mutants and evidence for clustering of men genes. J Bacteriol. 1981 Jan;145(1):321–327. doi: 10.1128/jb.145.1.321-327.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Tao X., Murphy J. R. Determination of the minimal essential nucleotide sequence for diphtheria tox repressor binding by in vitro affinity selection. Proc Natl Acad Sci U S A. 1994 Sep 27;91(20):9646–9650. doi: 10.1073/pnas.91.20.9646. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Zuber P., Losick R. Use of a lacZ fusion to study the role of the spoO genes of Bacillus subtilis in developmental regulation. Cell. 1983 Nov;35(1):275–283. doi: 10.1016/0092-8674(83)90230-1. [DOI] [PubMed] [Google Scholar]
  31. de Lorenzo V., Wee S., Herrero M., Neilands J. B. Operator sequences of the aerobactin operon of plasmid ColV-K30 binding the ferric uptake regulation (fur) repressor. J Bacteriol. 1987 Jun;169(6):2624–2630. doi: 10.1128/jb.169.6.2624-2630.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]

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