1887

Abstract

The surface (S)-layer gene region of the Gram-positive bacterium ATCC 14067 was identified on fosmid clones, sequenced and compared with the genome sequence of ATCC 13032, whose cell surface is devoid of an ordered S-layer lattice. A 5·97 kb DNA region that is absent from the ATCC 13032 chromosome was identified. This region includes , the structural gene encoding the S-layer protomer PS2, and six additional coding sequences. PCR experiments demonstrated that the respective DNA region is conserved in different wild-type strains capable of S-layer formation. The DNA region is flanked by a 7 bp direct repeat, suggesting that illegitimate recombination might be responsible for gene loss in ATCC 13032. Transfer of the cloned gene restored the PS2 phenotype of ATCC 13032, as confirmed by visualization of the PS2 proteins by SDS-PAGE and imaging of ordered hexagonal S-layer lattices on living cells by atomic force microscopy. Furthermore, the promoter of the gene was mapped by 5′ rapid amplification of cDNA ends PCR and the corresponding DNA fragment was used in DNA affinity purification assays. A 30 kDa protein specifically binding to the promoter region of the gene was purified. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry and peptide mass fingerprinting of the purified protein led to the identification of the putative transcriptional regulator Cg2831, belonging to the LuxR regulatory protein family. Disruption of the gene in resulted in an almost complete loss of PS2 synthesis. These results suggested that Cg2831 is a transcriptional activator of gene expression in .

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2006-04-01
2024-12-09
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References

  1. Altschul S. F, Madden T. L, Schaffer A. A, Zhang J, Zhang Z, Miller W, Lipman D. J. 1997; Gapped blast and psi-blast: a new generation of protein database search programs. Nucleic Acids Res 25:3389–3402 [CrossRef]
    [Google Scholar]
  2. Beveridge T. J, Pouwels P. H, Sara M. & 22 other authors; 1997; Functions of S-layers. FEMS Microbiol Rev 20:99–149 [CrossRef]
    [Google Scholar]
  3. Blaser M. J, Smith P. F, Hopkins J. A, Heinzer I, Bryner J. H, Wang W. L. 1987; Pathogenesis of Campylobacter fetus infections: serum resistance with high-molecular-weight surface proteins. J Infect Dis 155:696–706 [CrossRef]
    [Google Scholar]
  4. Bonamy C, Guyonvarch A, Reyes O, David F, Leblon G. 1990; Interspecies electro-transformation in Corynebacteria. FEMS Microbiol Lett 54:263–269
    [Google Scholar]
  5. Boos W, Shuman H. 1998; Maltose/maltodextrin system of Escherichia coli : transport, metabolism and regulation. Microbiol Mol Biol Rev 62:204–229
    [Google Scholar]
  6. Boot H. J, Pouwels P. H. 1996; Expression, secretion and antigenic variation of bacterial S-layer proteins. Mol Microbiol 21:1117–1123 [CrossRef]
    [Google Scholar]
  7. Brune I, Brinkrolf K, Kalinowski J, Pühler A, Tauch A. 2005; The individual and common repertoire of DNA-binding transcriptional regulators of Corynebacterium glutamicum , Corynebacterium efficiens , Corynebacterium diphtheriae and Corynebacterium jeikeium deduced from the complete genome sequences. BMC Genomics 6:86 [CrossRef]
    [Google Scholar]
  8. Capela D, Barloy-Hubler F, Gatius M.-T, Gouzy J, Galibert F. 1999; A high-density physical map of Sinorhizobium meliloti 1021 chromosome derived from bacterial artificial chromosome library. Proc Natl Acad Sci U S A 96:9357–9362 [CrossRef]
    [Google Scholar]
  9. Castan P, de Pedro M. A, Risco C, Valles C, Fernandez L. A, Schwarz H, Berenguer J. 2001; Multiple regulatory mechanisms act on the 5′ untranslated region of the S-layer gene from Thermus thermophilus HB8. J Bacteriol 183:1491–1494 [CrossRef]
    [Google Scholar]
  10. Chami M, Bayan N, Dedieu J.-C, Leblon G, Shechter E, Gulik-Krzywicki T. 1995; Organization of the outer layers of the cell envelope of Corynebacterium glutamicum : a combined freeze-etch electron microscopy and biochemical study. J Biol Cell 83:219–229 [CrossRef]
    [Google Scholar]
  11. Chami M, Bayan N, Peyret J. L, Gulik-Krzywicki T, Leblon G, Shechter E. 1997; The S-layer protein of Corynebacterium glutamicum is anchored to the cell wall by its C-terminal hydrophobic domain. Mol Microbiol 23:483–492 [CrossRef]
    [Google Scholar]
  12. Chu S, Trust T. J. 1993; An Aeromonas salmonicida gene which influences A-protein expression in Escherichia coli encodes a protein containing an ATP-binding cassette and maps beside the surface array protein gene. J Bacteriol 175:3105–3114
    [Google Scholar]
  13. Combet C, Blanchet C, Geourjon C, Deleage G. 2000; NPS@: network protein sequence analysis. Trends Biochem Sci 25:147–150 [CrossRef]
    [Google Scholar]
  14. Cui Y, Chatterjee A, Hasegawa H, Dixit V, Leigh N, Chatterjee A. K. 2005; ExpR, a LuxR homolog of Erwinia carotovora subsp. carotovora , activates transcription of rsmA , which specifies a global regulatory RNA-binding protein. J Bacteriol 187:4792–4803 [CrossRef]
    [Google Scholar]
  15. Cramer A, Gerstmeir R, Schaffer S, Bott M, Eikmanns B. J. 2006; Identification of RamA, a novel LuxR-type transcriptional regulator of genes involved in acetate metabolism of Corynebacterium glutamicum . J Bacteriol (in press)
    [Google Scholar]
  16. Eikmanns B. J, Thum-Schmitz N, Eggeling L, Ludtke K. U, Sahm H. 1994; Nucleotide sequence, expression and transcriptional analysis of the Corynebacterium glutamicum gltA gene encoding citrate synthase. Microbiology 140:1817–1828 [CrossRef]
    [Google Scholar]
  17. Farabaugh P. J, Schmeissner U, Hofer M, Miller J. H. 1978; Genetic studies of the Lac repressor. VII. On the molecular nature of spontaneous hotspots in the lacI gene of Escherichia coli . J Mol Biol 126:847–857 [CrossRef]
    [Google Scholar]
  18. Fernandez-Herrero L. A, Olabarria G, Berenguer J. 1997; Surface proteins and a novel transcription factor regulate the expression of the S-layer gene in Thermus thermophilus HB8. Mol Microbiol 24:61–72 [CrossRef]
    [Google Scholar]
  19. Fisher J. A, Smit J, Agabian N. 1988; Transcriptional analysis of the major surface array gene of Caulobacter crescentus . J Bacteriol 170:4706–4713
    [Google Scholar]
  20. Fujimoto S, Umeda A, Takade A, Murata K, Amako K. 1989; Hexagonal surface layer of Campylobacter fetus isolated from humans. Infect Immun 57:2563–2565
    [Google Scholar]
  21. Fujita M, Moriya T, Fujimoto S, Hara N, Amako K. 1997; A deletion in the sapA homologue cluster is responsible for the loss of the S-layer in Campylobacter fetus strain TK. Arch Microbiol 167:196–201 [CrossRef]
    [Google Scholar]
  22. Fuqua W. C, Winans S. C, Greenberg E. P. 1994; Quorum sensing in bacteria: the LuxR-LuxI family of cell density-response transcriptional regulators. J Bacteriol 176:269–275
    [Google Scholar]
  23. Garcia M. M, Lutz-Wallace C. L, Denes A. S, Eaglesome M. D, Holst E, Blaser M. J. 1995; Protein shift and antigenic variation in the S-layer of Campylobacter fetus subsp. venerealis during bovine infection accompanied by genomic rearrangement of sapA homologues. J Bacteriol 177:1976–1980
    [Google Scholar]
  24. 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 A 87:4645–4649 [CrossRef]
    [Google Scholar]
  25. Guvener Z. T, McCarter L. L. 2003; Multiple regulators control capsular polysaccharide production in Vibrio parahaemolyticus . J Bacteriol 185:5431–5441 [CrossRef]
    [Google Scholar]
  26. Hansmeier N, Bartels F. W, Ros R, Anselmetti D, Tauch A, Pühler A, Kalinowski J. 2004; Classification of hyper-variable Corynebacterium glutamicum surface-layer proteins by sequence analyses and atomic force microscopy. J Biotechnol 112:177–193 [CrossRef]
    [Google Scholar]
  27. Hermann T. 2003; Industrial production of amino acids by coryneform bacteria. J Biotechnol 104:155–172 [CrossRef]
    [Google Scholar]
  28. Ho Y.-S. J, Burden L. M, Hurley J. H. 2000; Structure of the GAF domain, a ubiquitous signaling motif and a new class of cyclic GMP receptor. EMBO J 19:5288–5299 [CrossRef]
    [Google Scholar]
  29. Houssin C, Nguyen D. T, Leblon G, Bayan N. 2002; S-layer protein transport across the cell wall of Corynebacterium glutamicum : in vivo kinetics and energy requirements. FEMS Microbiol Lett 217:71–79 [CrossRef]
    [Google Scholar]
  30. Kahala M, Sovijoki K, Palva A. 1997; In vivo expression of the Lactobacillus brevis S-layer gene. J Bacteriol 179:284–286
    [Google Scholar]
  31. Kalinowski J, Bathe B, Bartels D. & 24 other authors; 2003; The complete Corynebacterium glutamicum ATCC 13032 genome sequence and its impact on the production of l-aspartate-derived amino acids and vitamins. J Biotechnol 104:5–25 [CrossRef]
    [Google Scholar]
  32. Katsumata R, Ozaki A, Oka T, Furuya A. 1984; Protoplast transformation of glutamate-producing bacteria with plasmid DNA. J Bacteriol 159:306–311
    [Google Scholar]
  33. Krogh A, Larsson B, von Heijne G, Sonhammer E. L. 2001; Predicting transmembrane protein topology with a hidden Markov model: application to complete genomes. J Mol Biol 305:567–580 [CrossRef]
    [Google Scholar]
  34. Kumagai M, Ikeda H. 1991; Molecular analysis of the recombination junctions of λbio transducing phages. Mol Gen Genet 230:60–64 [CrossRef]
    [Google Scholar]
  35. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685 [CrossRef]
    [Google Scholar]
  36. Lopez P, Espinosa M, Greenberg B, Lacks S. A. 1984; Generation of deletions in pneumococcal mal genes cloned in Bacillus subtilis . Proc Natl Acad Sci U S A 81:5189–5193 [CrossRef]
    [Google Scholar]
  37. Madan Babu M, Teichmann S. A. 2003; Functional determinants of transcription factors in Escherichia coli : protein families and binding sites. Trends Genet 19:75–79 [CrossRef]
    [Google Scholar]
  38. Marchler-Bauer A, Anderson J. B, DeWeese-Sott C. 24 other authors 2003; CDD: a curated Entrez database of conserved domain alignments. Nucleic Acids Res 31:383–387 [CrossRef]
    [Google Scholar]
  39. Mateos L. M, Pisabarro A, Patek M, Malumbres M, Guerrero C, Eikmanns B. J, Sahm H, Martin J. F. 1994; Transcriptional analysis and regulatory signals of the hom - thrB cluster of Brevibacterium lactofermentum . J Bacteriol 176:7362–7371
    [Google Scholar]
  40. Mateos L. M, Schäfer A, Kalinowski J, Martin J. F, Pühler A. 1996; Integration of narrow-host-range vectors from Escherichia coli into the genomes of amino acid-producing cornyebacteria after intergenic conjugation. J Bacteriol 178:5768–5775
    [Google Scholar]
  41. Merino S, Alberti S, Tomas J. M. 1994; Aeromonas salmonicida resistance to complement-mediated killing. Infect Immun 62:5483–5490
    [Google Scholar]
  42. Messner P, Sleytr U. B. 1992; Crystalline bacterial cell-surface layers. Adv Microb Physiol 33:213–275
    [Google Scholar]
  43. Mignot T, Mesnage S, Couture-Tosi E, Mock M, Fouet A. 2002; Developmental switch of S-layer protein synthesis in Bacillus anthracis . Mol Microbiol 43:1615–1627 [CrossRef]
    [Google Scholar]
  44. Mignot T, Couture-Tosi E, Mesnage S, Mock M, Fouet A. 2004; In vivo Bacillus anthracis gene expression requires PagR as an intermediate effector of the AtxA signalling cascade. Int J Med Microbiol 293:619–624 [CrossRef]
    [Google Scholar]
  45. Noonan B, Trust T. J. 1995; Molecular analysis of an A-protein secretion mutant of Aeromonas salmonicida reveals a surface layer-specific protein secretion pathway. J Mol Biol 248:316–327
    [Google Scholar]
  46. Patek M, Eikmanns B. J, Patek K, Sahm H. 1996; Promoters from Corynebacterium glutamicum : cloning, molecular analysis and search for a consensus motif. Micobiology 142:1297–1309
    [Google Scholar]
  47. Perkins D. N, Pappin D. J, Creasy D. M, Cottrell J. S. 1999; Probability-based protein identification by searching sequence databases using mass spectrometry data. Electrophoresis 20:3551–3567 [CrossRef]
    [Google Scholar]
  48. Peyret J. L, Bayan N, Joliff G, Gulik-Krzywicki T, Mathieu L, Shechter E, Leblon G. 1993; Characterization of the cspB gene encoding PS2, an ordered surface-layer protein in Corynebacterium glutamicum . Mol Microbiol 9:97–109 [CrossRef]
    [Google Scholar]
  49. Pfaffl M. 2001; A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res 29:45 [CrossRef]
    [Google Scholar]
  50. 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 Biotechnol 103:51–63 [CrossRef]
    [Google Scholar]
  51. Sambrook J, Fritsch E. F, Maniatis T. 1989 Molecular Cloning: a Laboratory Manual, 2nd edn.. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  52. Sara M, Sleytr U. B. 2000; S-layer proteins. J Bacteriol 182:859–868 [CrossRef]
    [Google Scholar]
  53. Schäfer A, Tauch A, Jäger 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 . Gene 145:69–73 [CrossRef]
    [Google Scholar]
  54. Scheuring S, Stahlberg H, Chami M, Houssin C, Rigaud J.-L, Engel A. 2002; Charting and unzipping the surface layer of Corynebacterium glutamicum with the atomic force microscope. Mol Microbiol 44:675–684 [CrossRef]
    [Google Scholar]
  55. Schneitz C, Nuotio L, Lounatma K. 1993; Adhesion of Lactobacillus acidophilus to avian intestinal epithelial cells mediated by the crystalline bacterial cell surface layer (S-layer). J Appl Bacteriol 74:290–294 [CrossRef]
    [Google Scholar]
  56. Schweizer H. P. 1991; The agmR gene, an environmentally responsive gene, complements defective glpR , which encodes the putative activator for glycerol metabolism in Pseudomonas aeruginosa . J Bacteriol 173:6798–6806
    [Google Scholar]
  57. Sleytr U. B. 1997; I. Basic and applied S-layer research: an overview. FEMS Microbiol Rev 20:5–12 [CrossRef]
    [Google Scholar]
  58. Sleytr U. B, Messner P. 1983; Crystalline surface layers on bacteria. Annu Rev Microbiol 37:311–339 [CrossRef]
    [Google Scholar]
  59. Sleytr U. B, Sara M. 1997; Bacterial and archeal S-layer proteins: structure–function relationships and their biotechnological applications. Trends Biotechnol 15:20–26 [CrossRef]
    [Google Scholar]
  60. Soual-Hoebeke E, de Sousa-D'Auria C, Chami M, Baucher M. F, Guyonvarch A, Bayan N, Salim K, Leblon G. 1999; S-layer protein production by Corynebacterium strains is dependent on the carbon source. Microbiology 145:3399–3408
    [Google Scholar]
  61. Staden R. 1996; The Staden sequence analysis package. Mol Biotechnol 5:233–241 [CrossRef]
    [Google Scholar]
  62. Suzuki K, Wang X, Weilbacher T, Pernestig A. K, Melefors O, Georgellis D, Babitzke P, Romeo T. 2002; Regulatory circuitry of the CsrA/CrsB and BarA/UvrY systems in Escherichia coli . J Bacteriol 184:5130–5140 [CrossRef]
    [Google Scholar]
  63. Tummuru M. K. R, Blaser M. J. 1992; Characterization of the Campylobacter fetus sapA promoter: evidence that the sapA promoter is deleted in spontaneous mutant strains. J Bacteriol 174:5916–5922
    [Google Scholar]
  64. Tummuru M. K. R, Blaser M. J. 1993; Rearrangement of sapA homologues with conserved and variable regions in Campylobacter fetus . Proc Natl Acad Sci U S A 174:5916–5922
    [Google Scholar]
  65. Vidal-Ingigliardi D, Richet E, Raibaud O. 1991; Two MalT binding sites in direct repeat: a structural motif involved in the activation of all the promoters of the maltose regulons in Escherichia coli and Klebsiella pneumoniae . J Mol Biol 218:323–334 [CrossRef]
    [Google Scholar]
  66. Vidgren G, Palva I, Pakkanen R, Lounatmaa K, Palva A. 1992; S-layer gene of Lactobacillus brevis : cloning by polymerase chain reaction and determination of the nucleotide sequence. J Bacteriol 174:7419–7427
    [Google Scholar]
  67. Yanisch-Perron C, Vieira J, Messing J. 1985; Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene 33:103–119 [CrossRef]
    [Google Scholar]
  68. Yi T. M, Stearns D, Demple B. 1988; Illegitimate recombination in an Escherichia coli plasmid: modulation by DNA damage and a new bacterial gene. J Bacteriol 170:2898–2903
    [Google Scholar]
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