Skip to main content
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1996 Sep;178(18):5388–5394. doi: 10.1128/jb.178.18.5388-5394.1996

The Lactobacillus acidophilus S-layer protein gene expression site comprises two consensus promoter sequences, one of which directs transcription of stable mRNA.

H J Boot 1, C P Kolen 1, F J Andreadaki 1, R J Leer 1, P H Pouwels 1
PMCID: PMC178355  PMID: 8808926

Abstract

S-proteins are proteins which form a regular structure (S-layer) on the outside of the cell walls of many bacteria. Two S-protein-encoding genes are located in opposite directions on a 6.0-kb segment of the chromosome of Lactobacillus acidophilus ATCC 4356 bacteria. Inversion of this chromosomal segment occurs through recombination between two regions with identical sequences, thereby interchanging the expressed and the silent genes. In this study, we show that the region involved in recombination also has a function in efficient S-protein production. Two promoter sequences are present in the S-protein gene expression site, although only the most downstream promoter (P-1) is used to direct mRNA synthesis. S-protein mRNA directed by this promoter has a half-life of 15 min. Its untranslated leader can form a stable secondary structure in which the 5' end is base paired, whereas the ribosome-binding site is exposed. Truncation of this leader sequence results in a reduction in protein production, as shown by reporter gene analysis of Lactobacillus casei. The results obtained indicate that the untranslated leader sequence of S-protein mRNA is involved in efficient S-protein production.

Full Text

The Full Text of this article is available as a PDF (508.2 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Adachi T., Yamagata H., Tsukagoshi N., Udaka S. Multiple and tandemly arranged promoters of the cell wall protein gene operon in Bacillus brevis 47. J Bacteriol. 1989 Feb;171(2):1010–1016. doi: 10.1128/jb.171.2.1010-1016.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Adachi T., Yamagata H., Tsukagoshi N., Udaka S. Repression of the cell wall protein gene operon in Bacillus brevis 47 by magnesium and calcium ions. J Bacteriol. 1991 Jul;173(13):4243–4245. doi: 10.1128/jb.173.13.4243-4245.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bingle W. H., Doran J. L., Page W. J. Regular surface layer of Azotobacter vinelandii. J Bacteriol. 1984 Jul;159(1):251–259. doi: 10.1128/jb.159.1.251-259.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Boot H. J., Kolen C. P., Pouwels P. H. 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. J Bacteriol. 1995 Dec;177(24):7222–7230. doi: 10.1128/jb.177.24.7222-7230.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. 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]
  6. Bouvet P., Belasco J. G. Control of RNase E-mediated RNA degradation by 5'-terminal base pairing in E. coli. Nature. 1992 Dec 3;360(6403):488–491. doi: 10.1038/360488a0. [DOI] [PubMed] [Google Scholar]
  7. Brückner R. A series of shuttle vectors for Bacillus subtilis and Escherichia coli. Gene. 1992 Dec 1;122(1):187–192. doi: 10.1016/0378-1119(92)90048-t. [DOI] [PubMed] [Google Scholar]
  8. Chu S., Gustafson C. E., Feutrier J., Cavaignac S., Trust T. J. Transcriptional analysis of the Aeromonas salmonicida S-layer protein gene vapA. J Bacteriol. 1993 Dec;175(24):7968–7975. doi: 10.1128/jb.175.24.7968-7975.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Doig P., Emödy L., Trust T. J. Binding of laminin and fibronectin by the trypsin-resistant major structural domain of the crystalline virulence surface array protein of Aeromonas salmonicida. J Biol Chem. 1992 Jan 5;267(1):43–49. [PubMed] [Google Scholar]
  10. Fisher J. A., Smit J., Agabian N. Transcriptional analysis of the major surface array gene of Caulobacter crescentus. J Bacteriol. 1988 Oct;170(10):4706–4713. doi: 10.1128/jb.170.10.4706-4713.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Grivell L. A., Reijnders L., Borst P. The effect of temperature and ionic strength of the electrophoretic mobility of yeast mitochondrial RNA. Eur J Biochem. 1971 Mar 1;19(1):64–72. doi: 10.1111/j.1432-1033.1971.tb01288.x. [DOI] [PubMed] [Google Scholar]
  12. Horinouchi S., Weisblum B. Nucleotide sequence and functional map of pE194, a plasmid that specifies inducible resistance to macrolide, lincosamide, and streptogramin type B antibodies. J Bacteriol. 1982 May;150(2):804–814. doi: 10.1128/jb.150.2.804-814.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. 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]
  14. Kim S. F., Baek S. J., Pack M. Y. Cloning and nucleotide sequence of the Lactobacillus casei lactate dehydrogenase gene. Appl Environ Microbiol. 1991 Aug;57(8):2413–2417. doi: 10.1128/aem.57.8.2413-2417.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kitazawa H., Matsumura K., Itoh T., Yamaguchi T. Interferon induction in murine peritoneal macrophage by stimulation with Lactobacillus acidophilus. Microbiol Immunol. 1992;36(3):311–315. doi: 10.1111/j.1348-0421.1992.tb01668.x. [DOI] [PubMed] [Google Scholar]
  16. Koval S. F., Hynes S. H. Effect of paracrystalline protein surface layers on predation by Bdellovibrio bacteriovorus. J Bacteriol. 1991 Apr;173(7):2244–2249. doi: 10.1128/jb.173.7.2244-2249.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. 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]
  18. Ludwig W., Weizenegger M., Betzl D., Leidel E., Lenz T., Ludvigsen A., Möllenhoff D., Wenzig P., Schleifer K. H. Complete nucleotide sequences of seven eubacterial genes coding for the elongation factor Tu: functional, structural and phylogenetic evaluations. Arch Microbiol. 1990;153(3):241–247. doi: 10.1007/BF00249075. [DOI] [PubMed] [Google Scholar]
  19. Matern H. T., Klein J. R., Henrich B., Plapp R. Determination and comparison of Lactobacillus delbrueckii ssp. lactis DSM7290 promoter sequences. FEMS Microbiol Lett. 1994 Sep 15;122(1-2):121–128. doi: 10.1111/j.1574-6968.1994.tb07154.x. [DOI] [PubMed] [Google Scholar]
  20. Matuschek M., Burchhardt G., Sahm K., Bahl H. Pullulanase of Thermoanaerobacterium thermosulfurigenes EM1 (Clostridium thermosulfurogenes): molecular analysis of the gene, composite structure of the enzyme, and a common model for its attachment to the cell surface. J Bacteriol. 1994 Jun;176(11):3295–3302. doi: 10.1128/jb.176.11.3295-3302.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. 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]
  22. Noonan B., Trust T. J. The leucine zipper of Aeromonas salmonicida AbcA is required for the transcriptional activation of the P2 promoter of the surface-layer structural gene, vapA, in Escherichia coli. Mol Microbiol. 1995 Jul;17(2):379–386. doi: 10.1111/j.1365-2958.1995.mmi_17020379.x. [DOI] [PubMed] [Google Scholar]
  23. 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]
  24. Paula S. J., Duffey P. S., Abbott S. L., Kokka R. P., Oshiro L. S., Janda J. M., Shimada T., Sakazaki R. Surface properties of autoagglutinating mesophilic aeromonads. Infect Immun. 1988 Oct;56(10):2658–2665. doi: 10.1128/iai.56.10.2658-2665.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Petersen C. Control of functional mRNA stability in bacteria: multiple mechanisms of nucleolytic and non-nucleolytic inactivation. Mol Microbiol. 1992 Feb;6(3):277–282. doi: 10.1111/j.1365-2958.1992.tb01469.x. [DOI] [PubMed] [Google Scholar]
  26. Peyret J. L., Bayan N., Joliff G., Gulik-Krzywicki T., Mathieu L., Schechter E., Leblon G. Characterization of the cspB gene encoding PS2, an ordered surface-layer protein in Corynebacterium glutamicum. Mol Microbiol. 1993 Jul;9(1):97–109. doi: 10.1111/j.1365-2958.1993.tb01672.x. [DOI] [PubMed] [Google Scholar]
  27. 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]
  28. Pum D., Messner P., Sleytr U. B. Role of the S layer in morphogenesis and cell division of the archaebacterium Methanocorpusculum sinense. J Bacteriol. 1991 Nov;173(21):6865–6873. doi: 10.1128/jb.173.21.6865-6873.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Ross W., Gosink K. K., Salomon J., Igarashi K., Zou C., Ishihama A., Severinov K., Gourse R. L. A third recognition element in bacterial promoters: DNA binding by the alpha subunit of RNA polymerase. Science. 1993 Nov 26;262(5138):1407–1413. doi: 10.1126/science.8248780. [DOI] [PubMed] [Google Scholar]
  30. 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]
  31. Selker E., Yanofsky C. Nucleotide sequence of the trpC-trpB intercistronic region from Salmonella typhimurium. J Mol Biol. 1979 May 15;130(2):135–143. doi: 10.1016/0022-2836(79)90422-4. [DOI] [PubMed] [Google Scholar]
  32. Shaw W. V. Chloramphenicol acetyltransferase from chloramphenicol-resistant bacteria. Methods Enzymol. 1975;43:737–755. doi: 10.1016/0076-6879(75)43141-x. [DOI] [PubMed] [Google Scholar]
  33. Smith P. K., Krohn R. I., Hermanson G. T., Mallia A. K., Gartner F. H., Provenzano M. D., Fujimoto E. K., Goeke N. M., Olson B. J., Klenk D. C. Measurement of protein using bicinchoninic acid. Anal Biochem. 1985 Oct;150(1):76–85. doi: 10.1016/0003-2697(85)90442-7. [DOI] [PubMed] [Google Scholar]
  34. 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]
  35. Stefano J. E., Gralla J. D. Spacer mutations in the lac ps promoter. Proc Natl Acad Sci U S A. 1982 Feb;79(4):1069–1072. doi: 10.1073/pnas.79.4.1069. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Toba T., Virkola R., Westerlund B., Bjorkman Y., Sillanpaa J., Vartio T., Kalkkinen N., Korhonen T. K. A Collagen-Binding S-Layer Protein in Lactobacillus crispatus. Appl Environ Microbiol. 1995 Jul;61(7):2467–2471. doi: 10.1128/aem.61.7.2467-2471.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Van Delft J. H., Verbeek H. M., De Jong P. J., Schmidt D. S., Talens A., Bosch L. Control of the tRNA-tufB operon in Escherichia coli. 1. rRNA gene dosage effects and growth-rate-dependent regulation. Eur J Biochem. 1988 Aug 1;175(2):355–362. doi: 10.1111/j.1432-1033.1988.tb14204.x. [DOI] [PubMed] [Google Scholar]
  38. 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]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES