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. 1996 Jan;178(2):357–365. doi: 10.1128/jb.178.2.357-365.1996

slpM, a gene coding for an "S-layer-like array" overexpressed in S-layer mutants of Thermus thermophilus HB8.

G Olabarría 1, L A Fernández-Herrero 1, J L Carrascosa 1, J Berenguer 1
PMCID: PMC177665  PMID: 8550453

Abstract

S-layer deletion mutants of Thermus thermophilus HB8 overproduce a regular array which surrounds groups of several cells. Averages of two-dimensional projections revealed a detailed architecture similar in general morphology and unit cell dimensions to that of the S-layer but having a different mass distribution. The structural components of these "S-layer-like arrays" are a group of three proteins of 52 (P52), 50 (P50), and 36 (P36) kDa, which are overexpressed in S-layer mutants. These three proteins specifically bind antibodies against P52, suggesting that the smaller proteins correspond to fragments derived from P52. This hypothesis was demonstrated by the identity of the trypsin digestion products of P52 and P50. The gene slpM, responsible for the synthesis of P52, was cloned by using synthetic oligonucleotides designed from partial amino acid sequences of P52 and P50. When slpM was expressed in Escherichia coli, proteins specifically recognized by anti-P52 antiserum whose electrophoretic mobilities were similar to those of P52 and P36 were detected. The sequence of slpM revealed the existence of an open reading frame in which the amino termini of P52, P50, and P36 were identified. The unprocessed product of slpM is a 469-amino-acid-long polypeptide whose theoretical M(r) (52,131) was in good agreement with the electrophoretic mobility of P52. The properties deduced for the product of slpM are very different from those of any S-layer protein so far sequenced. The possible roles of SlpM in wild-type cells are discussed.

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Selected References

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  1. Castón J. R., Berenguer J., de Pedro M. A., Carrascosa J. L. S-layer protein from Thermus thermophilus HB8 assembles into porin-like structures. Mol Microbiol. 1993 Jul;9(1):65–75. doi: 10.1111/j.1365-2958.1993.tb01669.x. [DOI] [PubMed] [Google Scholar]
  2. Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Donate L. E., Herranz L., Secilla J. P., Carazo J. M., Fujisawa H., Carrascosa J. L. Bacteriophage T3 connector: three-dimensional structure and comparison with other viral head-tail connecting regions. J Mol Biol. 1988 May 5;201(1):91–100. doi: 10.1016/0022-2836(88)90441-x. [DOI] [PubMed] [Google Scholar]
  4. Faraldo M. L., de Pedro M. A., Berenguer J. Cloning and expression in Escherichia coli of the structural gene coding for the monomeric protein of the S layer of Thermus thermophilus HB8. J Bacteriol. 1991 Sep;173(17):5346–5351. doi: 10.1128/jb.173.17.5346-5351.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Faraldo M. M., de Pedro M. A., Berenguer J. Sequence of the S-layer gene of Thermus thermophilus HB8 and functionality of its promoter in Escherichia coli. J Bacteriol. 1992 Nov;174(22):7458–7462. doi: 10.1128/jb.174.22.7458-7462.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Fernández-Herrero L. A., Olabarría G., Castón J. R., Lasa I., Berenguer J. Horizontal transference of S-layer genes within Thermus thermophilus. J Bacteriol. 1995 Oct;177(19):5460–5466. doi: 10.1128/jb.177.19.5460-5466.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gruber K., Sleytr U. B. Localized insertion of new S-layer during growth of Bacillus stearothermophilus strains. Arch Microbiol. 1988;149(6):485–491. doi: 10.1007/BF00446749. [DOI] [PubMed] [Google Scholar]
  8. Koyama Y., Arikawa Y., Furukawa K. A plasmid vector for an extreme thermophile, Thermus thermophilus. FEMS Microbiol Lett. 1990 Oct;60(1-2):97–101. doi: 10.1016/0378-1097(90)90352-q. [DOI] [PubMed] [Google Scholar]
  9. Koyama Y., Hoshino T., Tomizuka N., Furukawa K. Genetic transformation of the extreme thermophile Thermus thermophilus and of other Thermus spp. J Bacteriol. 1986 Apr;166(1):338–340. doi: 10.1128/jb.166.1.338-340.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. LENNOX E. S. Transduction of linked genetic characters of the host by bacteriophage P1. Virology. 1955 Jul;1(2):190–206. doi: 10.1016/0042-6822(55)90016-7. [DOI] [PubMed] [Google Scholar]
  11. Laemmli U. K., Favre M. Maturation of the head of bacteriophage T4. I. DNA packaging events. J Mol Biol. 1973 Nov 15;80(4):575–599. doi: 10.1016/0022-2836(73)90198-8. [DOI] [PubMed] [Google Scholar]
  12. Lasa I., Castón J. R., Fernández-Herrero L. A., de Pedro M. A., Berenguer J. Insertional mutagenesis in the extreme thermophilic eubacteria Thermus thermophilus HB8. Mol Microbiol. 1992 Jun;6(11):1555–1564. doi: 10.1111/j.1365-2958.1992.tb00877.x. [DOI] [PubMed] [Google Scholar]
  13. Lasa I., de Grado M., de Pedro M. A., Berenguer J. Development of Thermus-Escherichia shuttle vectors and their use for expression of the Clostridium thermocellum celA gene in Thermus thermophilus. J Bacteriol. 1992 Oct;174(20):6424–6431. doi: 10.1128/jb.174.20.6424-6431.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Mather M. W., Fee J. A. Development of plasmid cloning vectors for Thermus thermophilus HB8: expression of a heterologous, plasmid-borne kanamycin nucleotidyltransferase gene. Appl Environ Microbiol. 1992 Jan;58(1):421–425. doi: 10.1128/aem.58.1.421-425.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. 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]
  16. Rosenfeld J., Capdevielle J., Guillemot J. C., Ferrara P. In-gel digestion of proteins for internal sequence analysis after one- or two-dimensional gel electrophoresis. Anal Biochem. 1992 May 15;203(1):173–179. doi: 10.1016/0003-2697(92)90061-b. [DOI] [PubMed] [Google Scholar]
  17. 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]
  18. Vieira J., Messing J. Production of single-stranded plasmid DNA. Methods Enzymol. 1987;153:3–11. doi: 10.1016/0076-6879(87)53044-0. [DOI] [PubMed] [Google Scholar]
  19. Woese C. R., Kandler O., Wheelis M. L. Towards a natural system of organisms: proposal for the domains Archaea, Bacteria, and Eucarya. Proc Natl Acad Sci U S A. 1990 Jun;87(12):4576–4579. doi: 10.1073/pnas.87.12.4576. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. 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]

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