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. 1995 Sep;177(17):4857–4864. doi: 10.1128/jb.177.17.4857-4864.1995

Molybdate and regulation of mod (molybdate transport), fdhF, and hyc (formate hydrogenlyase) operons in Escherichia coli.

J K Rosentel 1, F Healy 1, J A Maupin-Furlow 1, J H Lee 1, K T Shanmugam 1
PMCID: PMC177258  PMID: 7665461

Abstract

Escherichia coli mutants with defined mutations in specific mod genes that affect molybdate transport were isolated and analyzed for the effects of particular mutations on the regulation of the mod operon as well as the fdhF and hyc operons which code for the components of the formate hydrogenlyase (FHL) complex. phi (hyc'-'lacZ+) mod double mutants produced beta-galactosidase activity only when they were cultured in medium supplemented with molybdate. This requirement was specific for molybdate and was independent of the moa, mob, and moe gene products needed for molybdopterin guanine dinucleotide (MGD) synthesis, as well as Mog protein. The concentration of molybdate required for FHL production by mod mutants was dependent on medium composition. In low-sulfur medium, the amount of molybdate needed by mod mutants for the production of half-maximal FHL activity was increased approximately 20 times by the addition of 40 mM of sulfate, mod mutants growing in low-sulfur medium transported molybdate through the sulfate transport system, as seen by the requirement of the cysA gene product for this transport. In wild-type E. coli, the mod operon is expressed at very low levels, and a mod+ merodiploid E. coli carrying a modA-lacZ fusion produced less than 20 units of beta-galactosidase activity. This level was increased by over 175 times by a mutation in the modA, modB, or modC gene. The addition of molybdate to the growth medium of a mod mutant lowered phi (modA'-'lacZ+) expression. Repression of the mod operon was sensitive to molybdate but was insensitive to mutations in the MGD synthetic pathway. These physiological and genetic experiments show that molybdate can be transported by one of the following three anion transport system in E. coli: the native system, the sulfate transport system (cysTWA gene products), and an undefined transporter. Upon entering the cytoplasm, molybdate branches out to mod regulation, fdhF and hyc activation, and metabolic conversion, leading to MGD synthesis and active molybdoenzyme synthesis.

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Selected References

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  1. Bachmann B. J. Linkage map of Escherichia coli K-12, edition 7. Microbiol Rev. 1983 Jun;47(2):180–230. doi: 10.1128/mr.47.2.180-230.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Baker K. P., Boxer D. H. Regulation of the chlA locus of Escherichia coli K12: involvement of molybdenum cofactor. Mol Microbiol. 1991 Apr;5(4):901–907. doi: 10.1111/j.1365-2958.1991.tb00764.x. [DOI] [PubMed] [Google Scholar]
  3. Bremer E., Silhavy T. J., Weinstock G. M. Transposable lambda placMu bacteriophages for creating lacZ operon fusions and kanamycin resistance insertions in Escherichia coli. J Bacteriol. 1985 Jun;162(3):1092–1099. doi: 10.1128/jb.162.3.1092-1099.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Böhm R., Sauter M., Böck A. Nucleotide sequence and expression of an operon in Escherichia coli coding for formate hydrogenlyase components. Mol Microbiol. 1990 Feb;4(2):231–243. doi: 10.1111/j.1365-2958.1990.tb00590.x. [DOI] [PubMed] [Google Scholar]
  5. Dubourdieu M., Andrade E., Puig J. Molybdenum and chlorate resistant mutants in Escherichia coli K12. Biochem Biophys Res Commun. 1976 Jun 7;70(3):766–773. doi: 10.1016/0006-291x(76)90658-6. [DOI] [PubMed] [Google Scholar]
  6. Falciani F., Terao M., Goldwurm S., Ronchi A., Gatti A., Minoia C., Li Calzi M., Salmona M., Cazzaniga G., Garattini E. Molybdenum(VI) salts convert the xanthine oxidoreductase apoprotein into the active enzyme in mouse L929 fibroblastic cells. Biochem J. 1994 Feb 15;298(Pt 1):69–77. doi: 10.1042/bj2980069. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Glaser J. H., DeMoss J. A. Phenotypic restoration by molybdate of nitrate reductase activity in chlD mutants of Escherichia coli. J Bacteriol. 1971 Nov;108(2):854–860. doi: 10.1128/jb.108.2.854-860.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hemschemeier S., Grund M., Keuntje B., Eichenlaub R. Isolation of Escherichia coli mutants defective in uptake of molybdate. J Bacteriol. 1991 Oct;173(20):6499–6506. doi: 10.1128/jb.173.20.6499-6506.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hinton S. M., Dean D. Biogenesis of molybdenum cofactors. Crit Rev Microbiol. 1990;17(3):169–188. doi: 10.3109/10408419009105724. [DOI] [PubMed] [Google Scholar]
  10. Hopper S., Babst M., Schlensog V., Fischer H. M., Hennecke H., Böck A. Regulated expression in vitro of genes coding for formate hydrogenlyase components of Escherichia coli. J Biol Chem. 1994 Jul 29;269(30):19597–19604. [PubMed] [Google Scholar]
  11. Hryniewicz M. M., Kredich N. M. The cysP promoter of Salmonella typhimurium: characterization of two binding sites for CysB protein, studies of in vivo transcription initiation, and demonstration of the anti-inducer effects of thiosulfate. J Bacteriol. 1991 Sep;173(18):5876–5886. doi: 10.1128/jb.173.18.5876-5886.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hryniewicz M., Palucha A., Hulanicka M. D. Construction of cys:lac gene fusions in Escherichia coli and their use in the isolation of constitutive cysBc mutants. J Gen Microbiol. 1988 Mar;134(3):763–769. doi: 10.1099/00221287-134-3-763. [DOI] [PubMed] [Google Scholar]
  13. Iuchi S., Cameron D. C., Lin E. C. A second global regulator gene (arcB) mediating repression of enzymes in aerobic pathways of Escherichia coli. J Bacteriol. 1989 Feb;171(2):868–873. doi: 10.1128/jb.171.2.868-873.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Iuchi S., Furlong D., Lin E. C. Differentiation of arcA, arcB, and cpxA mutant phenotypes of Escherichia coli by sex pilus formation and enzyme regulation. J Bacteriol. 1989 May;171(5):2889–2893. doi: 10.1128/jb.171.5.2889-2893.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Jones H. M., Gunsalus R. P. Regulation of Escherichia coli fumarate reductase (frdABCD) operon expression by respiratory electron acceptors and the fnr gene product. J Bacteriol. 1987 Jul;169(7):3340–3349. doi: 10.1128/jb.169.7.3340-3349.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Lawson C. L., Carey J. Tandem binding in crystals of a trp repressor/operator half-site complex. Nature. 1993 Nov 11;366(6451):178–182. doi: 10.1038/366178a0. [DOI] [PubMed] [Google Scholar]
  17. Lee J. H., Patel P., Sankar P., Shanmugam K. T. Isolation and characterization of mutant strains of Escherichia coli altered in H2 metabolism. J Bacteriol. 1985 Apr;162(1):344–352. doi: 10.1128/jb.162.1.344-352.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Lee J. H., Wendt J. C., Shanmugam K. T. Identification of a new gene, molR, essential for utilization of molybdate by Escherichia coli. J Bacteriol. 1990 Apr;172(4):2079–2087. doi: 10.1128/jb.172.4.2079-2087.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Lindblow-Kull C., Kull F. J., Shrift A. Single transporter for sulfate, selenate, and selenite in Escherichia coli K-12. J Bacteriol. 1985 Sep;163(3):1267–1269. doi: 10.1128/jb.163.3.1267-1269.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Lopez Corcuera G., Bastidas M., Dubourdieu M. Molybdenum uptake in Escherichia coli K12. J Gen Microbiol. 1993 Aug;139(8):1869–1875. doi: 10.1099/00221287-139-8-1869. [DOI] [PubMed] [Google Scholar]
  21. Maupin-Furlow J. A., Rosentel J. K., Lee J. H., Deppenmeier U., Gunsalus R. P., Shanmugam K. T. Genetic analysis of the modABCD (molybdate transport) operon of Escherichia coli. J Bacteriol. 1995 Sep;177(17):4851–4856. doi: 10.1128/jb.177.17.4851-4856.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Maupin J. A., Shanmugam K. T. Genetic regulation of formate hydrogenlyase of Escherichia coli: role of the fhlA gene product as a transcriptional activator for a new regulatory gene, fhlB. J Bacteriol. 1990 Sep;172(9):4798–4806. doi: 10.1128/jb.172.9.4798-4806.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Menon N. K., Chatelus C. Y., Dervartanian M., Wendt J. C., Shanmugam K. T., Peck H. D., Jr, Przybyla A. E. Cloning, sequencing, and mutational analysis of the hyb operon encoding Escherichia coli hydrogenase 2. J Bacteriol. 1994 Jul;176(14):4416–4423. doi: 10.1128/jb.176.14.4416-4423.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Miller J. B., Scott D. J., Amy N. K. Molybdenum-sensitive transcriptional regulation of the chlD locus of Escherichia coli. J Bacteriol. 1987 May;169(5):1853–1860. doi: 10.1128/jb.169.5.1853-1860.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Ota N., Galsworthy P. R., Pardee A. B. Genetics of sulfate transport by Salmonella typhimurium. J Bacteriol. 1971 Mar;105(3):1053–1062. doi: 10.1128/jb.105.3.1053-1062.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Pascal M. C., Burini J. F., Ratouchniak J., Chippaux M. Regulation of the nitrate reductase operon: effect of mutations in chlA, B, D and E genes. Mol Gen Genet. 1982;188(1):103–106. doi: 10.1007/BF00333001. [DOI] [PubMed] [Google Scholar]
  27. Pecher A., Zinoni F., Jatisatienr C., Wirth R., Hennecke H., Böck A. On the redox control of synthesis of anaerobically induced enzymes in enterobacteriaceae. Arch Microbiol. 1983 Nov;136(2):131–136. doi: 10.1007/BF00404787. [DOI] [PubMed] [Google Scholar]
  28. Phillips S. E., Manfield I., Parsons I., Davidson B. E., Rafferty J. B., Somers W. S., Margarita D., Cohen G. N., Saint-Girons I., Stockley P. G. Cooperative tandem binding of met repressor of Escherichia coli. Nature. 1989 Oct 26;341(6244):711–715. doi: 10.1038/341711a0. [DOI] [PubMed] [Google Scholar]
  29. Phillips S. E., Stockley P. G. Similarity of met and trp repressors. Nature. 1994 Mar 10;368(6467):106–106. doi: 10.1038/368106a0. [DOI] [PubMed] [Google Scholar]
  30. Rajagopalan K. V., Johnson J. L. The pterin molybdenum cofactors. J Biol Chem. 1992 May 25;267(15):10199–10202. [PubMed] [Google Scholar]
  31. Schlensog V., Birkmann A., Böck A. Mutations in trans which affect the anaerobic expression of a formate dehydrogenase (fdhF) structural gene. Arch Microbiol. 1989;152(1):83–89. doi: 10.1007/BF00447016. [DOI] [PubMed] [Google Scholar]
  32. Scott D., Amy N. K. Molybdenum accumulation in chlD mutants of Escherichia coli. J Bacteriol. 1989 Mar;171(3):1284–1287. doi: 10.1128/jb.171.3.1284-1287.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Shanmugam K. T., Stewart V., Gunsalus R. P., Boxer D. H., Cole J. A., Chippaux M., DeMoss J. A., Giordano G., Lin E. C., Rajagopalan K. V. Proposed nomenclature for the genes involved in molybdenum metabolism in Escherichia coli and Salmonella typhimurium. Mol Microbiol. 1992 Nov;6(22):3452–3454. doi: 10.1111/j.1365-2958.1992.tb02215.x. [DOI] [PubMed] [Google Scholar]
  34. Sirko A., Hryniewicz M., Hulanicka D., Böck A. Sulfate and thiosulfate transport in Escherichia coli K-12: nucleotide sequence and expression of the cysTWAM gene cluster. J Bacteriol. 1990 Jun;172(6):3351–3357. doi: 10.1128/jb.172.6.3351-3357.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Sperl G. T., DeMoss J. A. chlD gene function in molybdate activation of nitrate reductase. J Bacteriol. 1975 Jun;122(3):1230–1238. doi: 10.1128/jb.122.3.1230-1238.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Spiro S., Guest J. R. FNR and its role in oxygen-regulated gene expression in Escherichia coli. FEMS Microbiol Rev. 1990 Aug;6(4):399–428. doi: 10.1111/j.1574-6968.1990.tb04109.x. [DOI] [PubMed] [Google Scholar]
  37. Stewart V., MacGregor C. H. Nitrate reductase in Escherichia coli K-12: involvement of chlC, chlE, and chlG loci. J Bacteriol. 1982 Aug;151(2):788–799. doi: 10.1128/jb.151.2.788-799.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Stewart V. Nitrate respiration in relation to facultative metabolism in enterobacteria. Microbiol Rev. 1988 Jun;52(2):190–232. doi: 10.1128/mr.52.2.190-232.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. WILSON L. G., BANDURSKI R. S. Enzymatic reactions involving sulfate, sulfite, selenate, and molybdate. J Biol Chem. 1958 Oct;233(4):975–981. [PubMed] [Google Scholar]
  40. Yerkes J. H., Casson L. P., Honkanen A. K., Walker G. C. Anaerobiosis induces expression of ant, a new Escherichia coli locus with a role in anaerobic electron transport. J Bacteriol. 1984 Apr;158(1):180–186. doi: 10.1128/jb.158.1.180-186.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]

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