1887

Abstract

A novel regulatory gene, which performs an essential function in sulfur metabolism, has been identified in and was designated (ysteine and ethionine regulator in ). The -disrupted strain (Δ) lost the ability to grow on minimal medium, and was identified as a methionine and cysteine double auxotroph. The mutant strain proved unable to convert cysteine to methionine (and vice versa), and lost the ability to assimilate and reduce sulfate to sulfide. In the Δ strain, the mRNAs of the methionine biosynthetic genes , and were significantly reduced, and the activities of the methionine biosynthetic enzymes cystathionine -synthase, -acetylhomoserine sulfhydrylase, and cystathionine -lyase were relatively low, thereby suggesting that the gene exerts a positive regulatory effect on methionine biosynthetic genes. In addition, with the exception of , reduced transcription levels of the sulfur-assimilatory genes and were noted in the -disrupted strain, which suggests that sulfur assimilation is also under the positive control of the gene. Furthermore, the expression of the gene itself was strongly induced via the addition of cysteine or methionine alone, but not the introduction of both amino acids together to the growth medium. In addition, the expression of the gene was enhanced in an -disrupted strain, which suggests that is under the negative control of McbR, which has been identified as a global regulator of sulfur metabolism. DNA binding of the purified CmaR protein to the promoter region of its target genes could be demonstrated . No metabolite effector was required for the protein to bind DNA. These results demonstrated that the gene of plays a role similar to but distinct from that of the functional homologue of .

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2009-06-01
2020-08-14
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References

  1. Adams A., Kaiser C.. 1998; Methods in Yeast Genetics: a Cold Spring Harbor Laboratory course manual , 1997 edn. Plainview, NY: Cold Spring Harbor Laboratory;
  2. Burguière P., Fert J., Guillouard I., Auger S., Danchin A., Martin-Verstraete I.. 2005; Regulation of the Bacillus subtilis ytmI operon, involved in sulfur metabolism. J Bacteriol187:6019–6030
    [Google Scholar]
  3. Choi W. W., Park S. D., Lee S. M., Kim H. B., Kim Y., Lee H. S.. 2009; The whcA gene plays a negative role in oxidative stress response of Corynebacterium glutamicum . FEMS Microbiol Lett290:32–38
    [Google Scholar]
  4. Even S., Burguière P., Auger S., Soutourina O., Danchin A., Martin-Verstraete I.. 2006; Global control of cysteine metabolism by CymR in Bacillus subtilis . J Bacteriol188:2184–2197
    [Google Scholar]
  5. Fernandez M., Kleerebezem M., Kuipers O. P., Siezen R. J., van Kranenburg R.. 2002; Regulation of the metC-cysK operon, involved in sulfur metabolism in Lactococcus lactis . J Bacteriol184:82–90
    [Google Scholar]
  6. Follettie M. T., Peoples O. P., Agoropoulou C., Sinskey A. J.. 1993; Gene structure and expression of the Corynebacterium flavum N13 ask-asd operon. J Bacteriol175:4096–4103
    [Google Scholar]
  7. Glanemann C., Loos A., Gorret N., Willis L. B., O'Brien X. M., Lessard P. A., Sinskey A. J.. 2003; Disparity between changes in mRNA abundance and enzyme activity in Corynebacterium glutamicum : implications for DNA microarray analysis. Appl Microbiol Biotechnol61:61–68
    [Google Scholar]
  8. Grant S. G., Jessee J., Bloom F. R., Hanahan D.. 1990; Differential plasmid rescue from transgenic mouse DNAs into Escherichia coli methylation–restriction mutants. Proc Natl Acad Sci U S A87:4645–4649
    [Google Scholar]
  9. Guillouard I., Auger S., Hullo M. F., Chetouani F., Danchin A., Martin-Verstraete I.. 2002; Identification of Bacillus subtilis CysL, a regulator of the cysJI operon, which encodes sulfite reductase. J Bacteriol184:4681–4689
    [Google Scholar]
  10. Haitani Y., Awano N., Yamazaki M., Wada M., Nakamori S., Takagi H.. 2006; Functional analysis of l-serine O -acetyltransferase from Corynebacterium glutamicum . FEMS Microbiol Lett255:156–163
    [Google Scholar]
  11. Hullo M. F., Auger S., Soutourina O., Barzu O., Yvon M., Danchin A., Martin-Verstraete I.. 2007; Conversion of methionine to cysteine in Bacillus subtilis and its regulation. J Bacteriol189:187–197
    [Google Scholar]
  12. Hwang B. J., Kim Y., Kim H. B., Hwang H. J., Kim J. H., Lee H. S.. 1999; Analysis of Corynebacterium glutamicum methionine biosynthetic pathway: isolation and analysis of metB encoding cystathionine gamma-synthase. Mol Cells9:300–308
    [Google Scholar]
  13. Hwang B. J., Yeom H. J., Kim Y., Lee H. S.. 2002; Corynebacterium glutamicum utilizes both transsulfuration and direct sulfhydrylation pathways for methionine biosynthesis. J Bacteriol184:1277–1286
    [Google Scholar]
  14. Iwanicka-Nowicka R., Hryniewicz M. M.. 1995; A new gene, cbl , encoding a member of the LysR family of transcriptional regulators belongs to Escherichia coli cys regulon. Gene166:11–17
    [Google Scholar]
  15. Iwanicka-Nowicka R., Zielak A., Cook A. M., Thomas M. S., Hryniewicz M. M.. 2007; Regulation of sulfur assimilation pathways in Burkholderia cenocepacia : identification of transcription factors CysB and SsuR and their role in control of target genes. J Bacteriol189:1675–1688
    [Google Scholar]
  16. Kim J. W., Kim H. J., Kim Y., Lee M. S., Lee H. S.. 2001; Properties of the Corynebacterium glutamicum metC gene encoding cystathionine β -lyase. Mol Cells11:220–225
    [Google Scholar]
  17. Kim H. J., Kim T. H., Kim Y., Lee H. S.. 2004; Identification and characterization of glxR , a gene involved in regulation of glyoxylate bypass in Corynebacterium glutamicum . J Bacteriol186:3453–3460
    [Google Scholar]
  18. Kim T. H., Kim H. J., Park J. S., Kim Y., Kim P., Lee H. S.. 2005; Functional analysis of sigH expression in Corynebacterium glutamicum . Biochem Biophys Res Commun331:1542–1547
    [Google Scholar]
  19. Koch D. J., Rückert C., Albersmeier A., Hüser A. T., Tauch A., Pühler A., Kalinowski J.. 2005a; The transcriptional regulator SsuR activates expression of the Corynebacterium glutamicum sulphonate utilization genes in the absence of sulphate. Mol Microbiol58:480–494
    [Google Scholar]
  20. Koch D. J., Rückert C., Rey D. A., Mix A., Pühler A., Kalinowski J.. 2005b; Role of the ssu and seu genes of Corynebacterium glutamicum ATCC 13032 in utilization of sulfonates and sulfonate esters as sulfur sources. Appl Environ Microbiol71:6104–6114
    [Google Scholar]
  21. Kouzuma A., Endoh T., Omori T., Nojiri H., Yamane H., Habe H.. 2008; Transcription factors CysB and SfnR constitute the hierarchical regulatory system for the sulfate starvation response in Pseudomonas putida . J Bacteriol190:4521–4531
    [Google Scholar]
  22. Kredich N. M.. others 1996; Biosynthesis of cysteine. In Escherichia coli and Salmonella: Cellular and Molecular Biology , 2nd edn. pp514–527 Edited by Neidhardt F. C.. Washington, DC: American Society for Microbiology;
  23. Lee H. S.. 2005; Sulfur metabolism and its regulation. In Handbook of Corynebacterium glutamicum pp351–376 Edited by Eggeling L., Bott M.. Boca Raton, FL: Taylor & Francis;
  24. Lee H. S., Hwang B. J.. 2003; Methionine biosynthesis and its regulation in Corynebacterium glutamicum : parallel pathways of transsulfuration and direct sulfhydrylation. Appl Microbiol Biotechnol62:459–467
    [Google Scholar]
  25. Liebl W.. 2006; Corynebacterium – Nonmedical. In The Prokaryotes: a Handbook on the Biology of Bacteria , 3rd edn. pp796–818 Edited by Dworkin M., Falkow S., Rosenberg E., Schleifer K.-H., Stackebrandt E.. New York: Springer;
  26. Link A. J., Phillips D., Church G. M.. 1997; Methods for generating precise deletions and insertions in the genome of wild-type Escherichia coli : application to open reading frame characterization. J Bacteriol179:6228–6237
    [Google Scholar]
  27. MacNeil D. J., Occi J. L., Gewain K. M., MacNeil T., Gibbons P. H., Ruby C. L., Danis S. J.. 1992; Complex organization of the Streptomyces avermitilis genes encoding the avermectin polyketide synthase. Gene115:119–125
    [Google Scholar]
  28. Moll I., Grill S., Gualerzi C. O., Blasi U.. 2002; Leaderless mRNAs in bacteria: surprises in ribosomal recruitment and translational control. Mol Microbiol43:239–246
    [Google Scholar]
  29. Ostrowski J., Kredich N. M.. 1989; Molecular characterization of the cysJIH promoters of Salmonella typhimurium and Escherichia coli : regulation by cysB protein and N -acetyl-l-serine. J Bacteriol171:130–140
    [Google Scholar]
  30. Ostrowski J., Kredich N. M.. 1990; In vitro interactions of CysB protein with the cysJIH promoter of Salmonella typhimurium : inhibitory effects of sulfide. J Bacteriol172:779–785
    [Google Scholar]
  31. Park S. D., Lee J. Y., Kim Y., Kim J. H., Lee H. S.. 1998; Isolation and analysis of metA , a methionine biosynthetic gene encoding homoserine acetyltransferase in Corynebacterium glutamicum . Mol Cells8:286–294
    [Google Scholar]
  32. Park S. D., Lee S. N., Park I. H., Choi J. S., Jeong W. K., Kim Y. H., Lee H. S.. 2004; Isolation and characterization of transcriptional elements from Corynebacterium glutamicum . J Microbiol Biotechnol14:789–795
    [Google Scholar]
  33. Park S. D., Lee J. Y., Sim S. Y., Kim Y., Lee H. S.. 2007; Characteristics of methionine production by an engineered Corynebacterium glutamicum strain. Metab Eng9:327–336
    [Google Scholar]
  34. Patek M., Nesvera J., Guyonvarch A., Reyes O., Leblon G.. 2003; Promoters of Corynebacterium glutamicum . J Biotechnol104:311–323
    [Google Scholar]
  35. Rey D. A., Pühler A., Kalinowski J.. 2003; The putative transcriptional repressor McbR, member of the TetR-family, is involved in the regulation of the metabolic network directing the synthesis of sulfur containing amino acids in Corynebacterium glutamicum . J Biotechnol103:51–65
    [Google Scholar]
  36. Rey D. A., Nentwich S. S., Koch D. J., Rückert C., Pühler A., Tauch A., Kalinowski J.. 2005; The McbR repressor modulated by the effector substance S -adenosylhomocysteine controls directly the transcription of a regulon involved in sulphur metabolism of Corynebacterium glutamicum ATCC 13032. Mol Microbiol56:871–887
    [Google Scholar]
  37. Rossol I., Pühler A.. 1992; The Corynebacterium glutamicum aecD gene encodes a C–S lyase with α , β -elimination activity that degrades aminoethylcysteine. J Bacteriol174:2968–2977
    [Google Scholar]
  38. Rückert C., Kalinowski J.. 2008; Sulfur metabolism in Corynebacterium glutamicum . In Corynebacteria : Genomics and Molecular Biology pp117–240 Edited by Burkowski A. Norwich, UK: Caister Academic Press;
    [Google Scholar]
  39. Rückert C., Pühler A., Kalinowski J.. 2003; Genome-wide analysis of the l-methionine biosynthetic pathway in Corynebacterium glutamicum by targeted gene deletion and homologous complementation. J Biotechnol104:213–228
    [Google Scholar]
  40. Rückert C., Koch D. J., Rey D. A., Albersmeier A., Mormann S., Pühler A., Kalinowski J.. 2005; Functional genomics and expression analysis of the Corynebacterium glutamicum fpr2 - cysIXHDNYZ gene cluster involved in assimilatory sulphate reduction. BMC Genomics6:121
    [Google Scholar]
  41. Rückert C., Milse J., Albersmeier A., Koch D. J., Pühler A., Kalinowski J.. 2008; The dual transcriptional regulator CysR in Corynebacterium glutamicum ATCC13032 controls a subset of genes of the McbR regulon in response to the availability of sulphide acceptor molecules. BMC Genomics9:483
    [Google Scholar]
  42. Schäfer A., Tauch A., Jager W., Kalinowski J., Thierbach G., Pühler A.. 1994; Small mobilizable multi-purpose cloning vectors derived from the Escherichia coli plasmids pK18 and pK19: selection of defined deletions in the chromosome of Corynebacterium glutamicum . Gene145:69–73
    [Google Scholar]
  43. Shaw W. V.. 1975; Chloramphenicol acetyltransferase from chloramphenicol-resistant bacteria. Methods Enzymol43:737–755
    [Google Scholar]
  44. Sperandio B., Polard P., Ehrlich D. S., Renault P., Guedon E.. 2005; Sulfur amino acid metabolism and its control in Lactococcus lactis IL1403. J Bacteriol187:3762–3778
    [Google Scholar]
  45. Suda M., Teramoto H., Imamiya T., Inui M., Yukawa H.. 2008; Transcriptional regulation of Corynebacterium glutamicum methionine biosynthesis genes in response to methionine supplementation under oxygen deprivation. Appl Microbiol Biotechnol81:505–513
    [Google Scholar]
  46. Tanous C., Soutourina O., Raynal B., Hullo M. F., Mervelet P., Gilles A. M., Noirot P., Danchin A., England P., Martin-Verstraete I.. 2008; The CymR regulator in complex with the enzyme CysK controls cysteine metabolism in Bacillus subtilis . J Biol Chem283:35551–35560
    [Google Scholar]
  47. van der Ploeg J. R., Iwanicka-Nowicka R., Kertesz M. A., Leisinger T., Hryniewicz M. M.. 1997; Involvement of CysB and Cbl regulatory proteins in expression of the tauABCD operon and other sulfate starvation-inducible genes in Escherichia coli . J Bacteriol179:7671–7678
    [Google Scholar]
  48. van der Ploeg J. R., Eichhorn E., Leisinger T.. 2001; Sulfonate–sulfur metabolism and its regulation in Escherichia coli . Arch Microbiol176:1–8
    [Google Scholar]
  49. Vermeij P., Kertesz M. A.. 1999; Pathways of assimilative sulfur metabolism in Pseudomonas putida . J Bacteriol181:5833–5837
    [Google Scholar]
  50. Wada M., Awano N., Haisa K., Takagi H., Nakamori S.. 2002; Purification, characterization and identification of cysteine desulfhydrase of Corynebacterium glutamicum , and its relationship to cysteine production. FEMS Microbiol Lett217:103–107
    [Google Scholar]
  51. Wada M., Awano N., Yamazawa H., Takagi H., Nakamori S.. 2004; Purification and characterization of O -acetylserine sulfhydrylase of Corynebacterium glutamicum . Biosci Biotechnol Biochem68:1581–1583
    [Google Scholar]
  52. Wheeler P. R., Coldham N. G., Keating L., Gordon S. V., Wooff E. E., Parish T., Hewinson R. G.. 2005; Functional demonstration of reverse transsulfuration in the Mycobacterium tuberculosis complex reveals that methionine is the preferred sulfur source for pathogenic mycobacteria. J Biol Chem280:8069–8078
    [Google Scholar]
  53. Yeom H. J., Hwang B. J., Lee M. S., Kim Y. H., Lee H. S.. 2004; Regulation of enzymes involved in methionine biosynthesis in Corynebacterium glutamicum . J Microbiol Biotechnol14:373–378
    [Google Scholar]
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