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. 1995 Dec;177(24):7222–7230. doi: 10.1128/jb.177.24.7222-7230.1995

Identification, cloning, and nucleotide sequence of a silent S-layer protein gene of Lactobacillus acidophilus ATCC 4356 which has extensive similarity with the S-layer protein gene of this species.

H J Boot 1, C P Kolen 1, P H Pouwels 1
PMCID: PMC177603  PMID: 8522531

Abstract

The bacterial S-layer forms a regular structure, composed of a monolayer of one (glyco)protein, on the surfaces of many prokaryotic species. S-layers are reported to fulfil different functions, such as attachment structures for extracellular enzymes and major virulence determinants for pathogenic species. Lactobacillus acidophilus ATCC 4356, which originates from the human pharynx, possesses such an S-layer. No function has yet been assigned to the S-layer of this species. Besides the structural gene (slpA) for the S-layer protein (S-protein) which constitutes this S-layer, we have identified a silent gene (slpB), which is almost identical to slpA in two regions. From the deduced amino acid sequence, it appears that the mature SB-protein (44,884 Da) is 53% similar to the SA-protein (43,636 Da) in the N-terminal and middle parts of the proteins. The C-terminal parts of the two proteins are identical except for one amino acid residue. The physical properties of the deduced S-proteins are virtually the same. Northern (RNA) blot analysis shows that only the slpA gene is expressed in wild-type cells, in line with the results from sequencing and primer extension analyses, which reveal that only the slpA gene harbors a promoter, which is located immediately upstream of the region where the two genes are identical. The occurrence of in vivo chromosomal recombination between the two S-protein-encoding genes will be described elsewhere.

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Selected References

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  1. Belland R. J., Trust T. J. Cloning of the gene for the surface array protein of Aeromonas salmonicida and evidence linking loss of expression with genetic deletion. J Bacteriol. 1987 Sep;169(9):4086–4091. doi: 10.1128/jb.169.9.4086-4091.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Blaser M. J., Wang E., Tummuru M. K., Washburn R., Fujimoto S., Labigne A. High-frequency S-layer protein variation in Campylobacter fetus revealed by sapA mutagenesis. Mol Microbiol. 1994 Nov;14(3):453–462. doi: 10.1111/j.1365-2958.1994.tb02180.x. [DOI] [PubMed] [Google Scholar]
  3. Boot H. J., Kolen C. P., van Noort J. M., Pouwels P. H. S-layer protein of Lactobacillus acidophilus ATCC 4356: purification, expression in Escherichia coli, and nucleotide sequence of the corresponding gene. J Bacteriol. 1993 Oct;175(19):6089–6096. doi: 10.1128/jb.175.19.6089-6096.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bowditch R. D., Baumann P., Yousten A. A. Cloning and sequencing of the gene encoding a 125-kilodalton surface-layer protein from Bacillus sphaericus 2362 and of a related cryptic gene. J Bacteriol. 1989 Aug;171(8):4178–4188. doi: 10.1128/jb.171.8.4178-4188.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Catalanotti P., Rossano F., de Paolis P., Baroni A., Buttini G., Tufano M. A. Effects of Cetyltrimethylammonium naproxenate on the adherence of Gardnerella vaginalis, Mobiluncus curtisii, and Lactobacillus acidophilus to vaginal epithelial cells. Sex Transm Dis. 1994 Nov-Dec;21(6):338–344. doi: 10.1097/00007435-199411000-00008. [DOI] [PubMed] [Google Scholar]
  6. Chu S., Cavaignac S., Feutrier J., Phipps B. M., Kostrzynska M., Kay W. W., Trust T. J. Structure of the tetragonal surface virulence array protein and gene of Aeromonas salmonicida. J Biol Chem. 1991 Aug 15;266(23):15258–15265. [PubMed] [Google Scholar]
  7. Coconnier M. H., Bernet M. F., Kernéis S., Chauvière G., Fourniat J., Servin A. L. Inhibition of adhesion of enteroinvasive pathogens to human intestinal Caco-2 cells by Lactobacillus acidophilus strain LB decreases bacterial invasion. FEMS Microbiol Lett. 1993 Jul 1;110(3):299–305. doi: 10.1111/j.1574-6968.1993.tb06339.x. [DOI] [PubMed] [Google Scholar]
  8. Dworkin J., Tummuru M. K., Blaser M. J. A lipopolysaccharide-binding domain of the Campylobacter fetus S-layer protein resides within the conserved N terminus of a family of silent and divergent homologs. J Bacteriol. 1995 Apr;177(7):1734–1741. doi: 10.1128/jb.177.7.1734-1741.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Egelseer E., Schocher I., Sára M., Sleytr U. B. The S-layer from Bacillus stearothermophilus DSM 2358 functions as an adhesion site for a high-molecular-weight amylase. J Bacteriol. 1995 Mar;177(6):1444–1451. doi: 10.1128/jb.177.6.1444-1451.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Ishiguro E. E., Kay W. W., Ainsworth T., Chamberlain J. B., Austen R. A., Buckley J. T., Trust T. J. Loss of virulence during culture of Aeromonas salmonicida at high temperature. J Bacteriol. 1981 Oct;148(1):333–340. doi: 10.1128/jb.148.1.333-340.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Klaenhammer T. R. Genetics of bacteriocins produced by lactic acid bacteria. FEMS Microbiol Rev. 1993 Sep;12(1-3):39–85. doi: 10.1111/j.1574-6976.1993.tb00012.x. [DOI] [PubMed] [Google Scholar]
  12. Koval S. F., Murray R. The superficial protein arrays on bacteria. Microbiol Sci. 1986 Dec;3(12):357–361. [PubMed] [Google Scholar]
  13. Kuen B., Sleytr U. B., Lubitz W. Sequence analysis of the sbsA gene encoding the 130-kDa surface-layer protein of Bacillus stearothermophilus strain PV72. Gene. 1994 Jul 22;145(1):115–120. doi: 10.1016/0378-1119(94)90332-8. [DOI] [PubMed] [Google Scholar]
  14. Leblond P., Francou F. X., Simonet J. M., Decaris B. Pulsed-field gel electrophoresis analysis of the genome of Streptomyces ambofaciens strains. FEMS Microbiol Lett. 1990 Oct;60(1-2):79–88. doi: 10.1016/0378-1097(90)90349-u. [DOI] [PubMed] [Google Scholar]
  15. Link-Amster H., Rochat F., Saudan K. Y., Mignot O., Aeschlimann J. M. Modulation of a specific humoral immune response and changes in intestinal flora mediated through fermented milk intake. FEMS Immunol Med Microbiol. 1994 Nov;10(1):55–63. doi: 10.1111/j.1574-695X.1994.tb00011.x. [DOI] [PubMed] [Google Scholar]
  16. Lupas A., Engelhardt H., Peters J., Santarius U., Volker S., Baumeister W. Domain structure of the Acetogenium kivui surface layer revealed by electron crystallography and sequence analysis. J Bacteriol. 1994 Mar;176(5):1224–1233. doi: 10.1128/jb.176.5.1224-1233.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. McGroarty J. A. Probiotic use of lactobacilli in the human female urogenital tract. FEMS Immunol Med Microbiol. 1993 Apr;6(4):251–264. doi: 10.1111/j.1574-695X.1993.tb00337.x. [DOI] [PubMed] [Google Scholar]
  18. Messner P., Sleytr U. B. Crystalline bacterial cell-surface layers. Adv Microb Physiol. 1992;33:213–275. doi: 10.1016/s0065-2911(08)60218-0. [DOI] [PubMed] [Google Scholar]
  19. Orrhage K., Sillerström E., Gustafsson J. A., Nord C. E., Rafter J. Binding of mutagenic heterocyclic amines by intestinal and lactic acid bacteria. Mutat Res. 1994 Dec 1;311(2):239–248. doi: 10.1016/0027-5107(94)90182-1. [DOI] [PubMed] [Google Scholar]
  20. Pouwels P. H., Leer R. J. Genetics of lactobacilli: plasmids and gene expression. Antonie Van Leeuwenhoek. 1993;64(2):85–107. doi: 10.1007/BF00873020. [DOI] [PubMed] [Google Scholar]
  21. Pouwels P. H., Leunissen J. A. Divergence in codon usage of Lactobacillus species. Nucleic Acids Res. 1994 Mar 25;22(6):929–936. doi: 10.1093/nar/22.6.929. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Roussel Y., Colmin C., Simonet J. M., Decaris B. Strain characterization, genome size and plasmid content in the Lactobacillus acidophilus group (Hansen and Mocquot). J Appl Bacteriol. 1993 May;74(5):549–556. [PubMed] [Google Scholar]
  23. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Simonen M., Palva I. Protein secretion in Bacillus species. Microbiol Rev. 1993 Mar;57(1):109–137. doi: 10.1128/mr.57.1.109-137.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  26. Sára M., Pum D., Küpcü S., Messner P., Sleytr U. B. Isolation of two physiologically induced variant strains of Bacillus stearothermophilus NRS 2004/3a and characterization of their S-layer lattices. J Bacteriol. 1994 Feb;176(3):848–860. doi: 10.1128/jb.176.3.848-860.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Sára M., Sleytr U. B. Comparative studies of S-layer proteins from Bacillus stearothermophilus strains expressed during growth in continuous culture under oxygen-limited and non-oxygen-limited conditions. J Bacteriol. 1994 Dec;176(23):7182–7189. doi: 10.1128/jb.176.23.7182-7189.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Tummuru M. K., Blaser M. J. Rearrangement of sapA homologs with conserved and variable regions in Campylobacter fetus. Proc Natl Acad Sci U S A. 1993 Aug 1;90(15):7265–7269. doi: 10.1073/pnas.90.15.7265. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Vidgrén G., Palva I., Pakkanen R., Lounatmaa K., Palva A. S-layer protein gene of Lactobacillus brevis: cloning by polymerase chain reaction and determination of the nucleotide sequence. J Bacteriol. 1992 Nov;174(22):7419–7427. doi: 10.1128/jb.174.22.7419-7427.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Wang E., Garcia M. M., Blake M. S., Pei Z., Blaser M. J. Shift in S-layer protein expression responsible for antigenic variation in Campylobacter fetus. J Bacteriol. 1993 Aug;175(16):4979–4984. doi: 10.1128/jb.175.16.4979-4984.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Wildhaber I., Baumeister W. The cell envelope of Thermoproteus tenax: three-dimensional structure of the surface layer and its role in shape maintenance. EMBO J. 1987 May;6(5):1475–1480. doi: 10.1002/j.1460-2075.1987.tb02389.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Yamagata H., Adachi T., Tsuboi A., Takao M., Sasaki T., Tsukagoshi N., Udaka S. Cloning and characterization of the 5' region of the cell wall protein gene operon in Bacillus brevis 47. J Bacteriol. 1987 Mar;169(3):1239–1245. doi: 10.1128/jb.169.3.1239-1245.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]
  34. Zheng H. Y., Alcorn T. M., Cohen M. S. Effects of H2O2-producing lactobacilli on Neisseria gonorrhoeae growth and catalase activity. J Infect Dis. 1994 Nov;170(5):1209–1215. doi: 10.1093/infdis/170.5.1209. [DOI] [PubMed] [Google Scholar]
  35. von Heijne G. A new method for predicting signal sequence cleavage sites. Nucleic Acids Res. 1986 Jun 11;14(11):4683–4690. doi: 10.1093/nar/14.11.4683. [DOI] [PMC free article] [PubMed] [Google Scholar]

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