1887

Abstract

The cell surface of ATCC 12980 is completely covered with an oblique S-layer lattice. To investigate sequence identities and a common structure–function relationship in S-layer proteins of different wild-type strains, the nucleotide sequence encoding theS-layer protein SbsC of . ATCC 12980 was determined by PCR techniques. The entire sequence showed an ORF of 3297 bp predicted to encode a protein of 1099 aa with a theoretical molecular mass of 115409 Da and an isoelectric point of 573. Primer extension analysis suggested the existence of two promoter regions. Amino acid sequence comparison between SbsC and SbsA, a previously characterized S-layer protein of PV72/p6 which assembles into a hexagonally ordered lattice, revealed an identical secretion signal peptide, 85% identity for theN-terminal regions (aa 31–270) which do not carry any S-layer homologous motifs, but only 21% identity for the rest of the sequences. Affinity studies demonstrated that the N-terminal part of SbsC is necessary for recognition of a secondary cell wall polymer. This was in accordance with results obtained in a previous study for SbsA, thus confirming a common functional principle for the N-terminal parts of both S-layer proteins. The coding region cloned into the pET3a vector without its own upstream region, the signal sequence and the 3′ transcriptional terminator led to stable expression in .

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2000-02-01
2020-01-28
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References

  1. Adachi T., Yamagata H., Tsukagoshi N., Udaka S.. 1989; Multiple and tandemly arranged promoters of the cell wall protein gene operon in Bacillus brevis 47. J Bacteriol171:1010–1016
    [Google Scholar]
  2. Altschul S. F., Madden T. L., Schäffer 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 Res25:3389–3402[CrossRef]
    [Google Scholar]
  3. Boot H. J., Kolen C. P. A. M., Andreadaki F. J., Leer R. J., Pouwels P. H.. 1996; The Lactobacillus acidophilus S-layer protein gene expression site comprises two consensus promoter sequences, one of which directs transcription of stable mRNA. J Bacteriol178:5388–5394
    [Google Scholar]
  4. Brechtel E., Bahl H.. 1999; In Thermoanaerobacterium thermosulfurigenes EM1 S-layer homology domains do not attach to peptidoglycan. J Bacteriol181:5017–5023
    [Google Scholar]
  5. Chauvaux S., Matuschek M., Béguin P.. 1999; Distinct affinity of binding sites for S-layer homologous domains in Clostridium thermocellum and Bacillus anthracis. J Bacteriol181:2455–2458
    [Google Scholar]
  6. Don R. H., Cox P. T., Wainwright B. J., Baker K., Mattick J. S.. 1991; ‘‘Touchdown’’ PCR to circumvent spurious priming during gene amplification. Nucleic Acids Res19:4008[CrossRef]
    [Google Scholar]
  7. Egelseer E. M., Schocher I., Sleytr U. B., Sára M.. 1996; Evidence that an N-terminal S-layer protein fragment triggers the release of a cell associated high-molecular-weight amylase from Bacillus stearothermophilus ATCC 12980. J Bacteriol178:5602–5609
    [Google Scholar]
  8. Egelseer E. M., Leitner K., Jarosch M., Hotzy C., Zayni S., Sleytr U. B., Sára M.. 1998; The S-layer proteins of two Bacillus stearothermophilus wild-type strains are bound via their N-terminal region to a secondary cell wall polymer of identical chemical composition. J Bacteriol180:1488–1495
    [Google Scholar]
  9. Engelhardt H., Peters J.. 1998; Structural research on surface layers: a focus on stability, surface layer homology domains, and surface layer-cell wall interactions. J Struct Biol124:276–302[CrossRef]
    [Google Scholar]
  10. Etienne-Toumelin I., Sirard J.-C., Duflot E., Mock M., Fouet A.. 1995; Characterization of the Bacillus anthracis S-layer: cloning and sequencing of the structural gene. J Bacteriol177:614–620
    [Google Scholar]
  11. Ilk N., Kosma P., Puchberger M., Egelseer E. M., Mayer H. F., Sleytr U. B., Sára M.. 1999; Structural and functional analysis of the secondary cell wall polymer of Bacillus sphaericus CCM 2177 serving as an S-layer specific anchor. J Bacteriol181: (in press)
    [Google Scholar]
  12. Kahala M., Savijoki K., Palva A.. 1997; In vivo expression of the Lactobacillus brevis S-layer gene. J Bacteriol179:284–286
    [Google Scholar]
  13. Kuen B., Sleytr U. B., Lubitz W.. 1994; Sequence analysis of the sbsA gene encoding the 130-kDa surface-layer protein of Bacillus stearothermophilus PV72. Gene45:115–120
    [Google Scholar]
  14. Kuen B., Sára M., Lubitz W.. 1995; Heterologous expression and self-assembly of the S-layer protein SbsA of Bacillus stearothermophilus in Escherichia coli. Mol Microbiol19:495–503
    [Google Scholar]
  15. Kuen B., Koch A., Asenbauer E., Sára M., Lubitz W.. 1997; Molecular characterization of the second S-layer gene sbsB of Bacillus stearothermophilus PV72 expressed by oxidative stress. J Bacteriol179:1664–1670
    [Google Scholar]
  16. Laemmli U. K.. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature227:680–685[CrossRef]
    [Google Scholar]
  17. Lupas A.. 1996; A circular permutation event in the evolution of the SLH-domain?. Mol Microbiol20:897–898[CrossRef]
    [Google Scholar]
  18. Lupas A., Engelhardt H., Peters J., Santarius U., Volker S., Baumeister W.. 1994; Domain structure of the Acetogenium kivui surface layer revealed by electron crystallography and sequence analysis. J Bacteriol176:1224–1233
    [Google Scholar]
  19. Mesnage S., Tosi-Couture E., Mock M., Gounon P., Fouet A.. 1997; Molecular characterization of the Bacillus anthracis main S-layer component: evidence that it is the major cell-associated antigen. Mol Microbiol23:1147–1155[CrossRef]
    [Google Scholar]
  20. Mesnage S., Tosi-Couture E., Fouet A.. 1999; Production and cell surface anchoring of functional fusions between the SLH motifs of the Bacillus anthracis S-layer proteins and the Bacillus subtilis levansucrase. Mol Microbiol31:927–936[CrossRef]
    [Google Scholar]
  21. Messner P., Hollaus F., Sleytr U. B.. 1984; Paracrystalline cell wall surface layers of different Bacillus stearothermophilus strains. Int J Syst Bacteriol34:202–210[CrossRef]
    [Google Scholar]
  22. Rost B., Sander C.. 1993; Prediction of protein structure at better than 70% accuracy. J Mol Biol232:584–599[CrossRef]
    [Google Scholar]
  23. 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]
  24. Sanger F., Nicklen S., Coulson A. R.. 1977; DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA74:5463–5467[CrossRef]
    [Google Scholar]
  25. Sára M., Sleytr U. B.. 1996; Crystalline bacterial cell surface layers (S-layers): from cell structure to biomimetics. Prog Biophys Mol Biol65:83–111
    [Google Scholar]
  26. Sára M., Kuen B., Mayer H. F., Mandl F., Schuster K. C., Sleytr U. B.. 1996; Dynamics in oxygen-induced changes in S-layer protein synthesis from Bacillus stearothermophilus PV72 and its S-layer deficient variant T5 in continuous culture and studies on the cell wall composition. J Bacteriol178:2108–2117
    [Google Scholar]
  27. Schäffer C., Kählig H., Christian R., Schulz G., Zayni S., Messner P.. 1999; The diacetamidodideoxyuronic-acid-containing glycan chain of Bacillus stearothermophilus NRS 2004/3a represents the secondary cell-wall polymer of wild-type B. stearothermophilus strains. Microbiology145:1575–1583[CrossRef]
    [Google Scholar]
  28. Scholz H., Kuen B., Lubitz W., Sára M.. 1997; S-layer variation in Bacillus stearothermophilus PV72. FEMS Microbiol Rev20:69–78
    [Google Scholar]
  29. Sleytr U. B., Messner P.. 1983; Crystalline surface layers on bacteria. Annu Rev Microbiol37:311–339[CrossRef]
    [Google Scholar]
  30. Sleytr U. B., Messner P., Pum D., Sára M.. 1993; Crystalline bacterial cell surface layers. Mol Microbiol10:911–916[CrossRef]
    [Google Scholar]
  31. Sleytr U. B., Messner P., Pum D., Sára M.. 1999; Crystalline bacterial cell surface layers (S-layers): from supramolecular cell structure to biomimetics and nanotechnology. Angew Chem Int Ed38:1034–1054[CrossRef]
    [Google Scholar]
  32. Studier F. W., Rosenberg A. H., Dunn J. J., Dubendorff J. W.. 1990; Use of T7 RNA polymerase to direct expression of cloned genes. Methods Enzymol185:60–89
    [Google Scholar]
  33. Transue T. R., Smith K. A., Mo H., Goldstein I. J., Saper M. A.. 1997; Structure of benzyl-T-antigen disaccharide bound to Amaranthus caudatus agglutinin. Nat Struct Biol4:779–783[CrossRef]
    [Google Scholar]
  34. Weis W. I.. 1997; Cell-surface carbohydrate recognition by animal and viral lectins. Curr Opin Struct Biol7:624–630[CrossRef]
    [Google Scholar]
  35. Zhu X., McVeigh R. R., Malathi P., Gosh B. K.. 1996; The complete nucleotide sequence of the Bacillus licheniformis NM105 S-layer encoding gene. Gene173:189–194[CrossRef]
    [Google Scholar]
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