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. 1995 Jul;177(14):4105–4112. doi: 10.1128/jb.177.14.4105-4112.1995

The ftsH gene of Bacillus subtilis is transiently induced after osmotic and temperature upshift.

E Deuerling 1, B Paeslack 1, W Schumann 1
PMCID: PMC177143  PMID: 7608085

Abstract

The ftsH gene of Bacillus subtilis has been identified as a salt-sensitive insertion mutation in strain UG1. Here, we show that UG1 has an insertion near the 3' end of ftsH. The salt sensitivity of this mutant was caused by reduction of ftsH mRNA levels by the synthesis of an artificial antisense RNA originating at a promoter located within the insertion and reading backwards into the ftsH gene. The salt-sensitive phenotype could be overcome by deleting the promoter from which the antisense RNA was transcribed. A physiological analysis of the isogenic wild-type strain in minimal medium revealed unimpaired growth at up to 1 M NaCl, and growth above 1.2 M NaCl was observed only after addition of the osmoprotectant proline or glycine betaine. In contrast, growth of strain UG1 was reduced at a salt concentration above 0.2 M, which could be rescued by the two compatible solutes already mentioned and also by trehalose. Primer extension revealed one potential transcription start site downstream of a putative vegetative promoter, which was activated after osmotic or temperature upshift. Northern (RNA blot) experiments led to the detection of a 2.1-kb transcript, suggesting that ftsH is monocistronic. A transcriptional fusion between ftsH and the gus reporter gene exhibited a twofold increase in beta-glucuronidase activity after osmotic upshift. To further confirm the need for an enhanced level of FtsH protein after osmotic upshift, the promoter was replaced by the sucrose-inducible promoter PsacB. Whereas this mutant strain could grow in the absence of inducer in LB medium, it stopped growth immediately after addition of 1.1 M NaCl. We conclude that an increased amount of FtsH protein is essential for B. subtilis to cope with an increase in osmolarity or temperature.

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Selected References

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  1. Akiyama Y., Ogura T., Ito K. Involvement of FtsH in protein assembly into and through the membrane. I. Mutations that reduce retention efficiency of a cytoplasmic reporter. J Biol Chem. 1994 Feb 18;269(7):5218–5224. [PubMed] [Google Scholar]
  2. Akiyama Y., Shirai Y., Ito K. Involvement of FtsH in protein assembly into and through the membrane. II. Dominant mutations affecting FtsH functions. J Biol Chem. 1994 Feb 18;269(7):5225–5229. [PubMed] [Google Scholar]
  3. Back J. F., Oakenfull D., Smith M. B. Increased thermal stability of proteins in the presence of sugars and polyols. Biochemistry. 1979 Nov 13;18(23):5191–5196. doi: 10.1021/bi00590a025. [DOI] [PubMed] [Google Scholar]
  4. Begg K. J., Tomoyasu T., Donachie W. D., Khattar M., Niki H., Yamanaka K., Hiraga S., Ogura T. Escherichia coli mutant Y16 is a double mutant carrying thermosensitive ftsH and ftsI mutations. J Bacteriol. 1992 Apr;174(7):2416–2417. doi: 10.1128/jb.174.7.2416-2417.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Boch J., Kempf B., Bremer E. Osmoregulation in Bacillus subtilis: synthesis of the osmoprotectant glycine betaine from exogenously provided choline. J Bacteriol. 1994 Sep;176(17):5364–5371. doi: 10.1128/jb.176.17.5364-5371.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chang A. C., Cohen S. N. Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid. J Bacteriol. 1978 Jun;134(3):1141–1156. doi: 10.1128/jb.134.3.1141-1156.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Csonka L. N., Hanson A. D. Prokaryotic osmoregulation: genetics and physiology. Annu Rev Microbiol. 1991;45:569–606. doi: 10.1146/annurev.mi.45.100191.003033. [DOI] [PubMed] [Google Scholar]
  8. Csonka L. N. Physiological and genetic responses of bacteria to osmotic stress. Microbiol Rev. 1989 Mar;53(1):121–147. doi: 10.1128/mr.53.1.121-147.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Ferrari F. A., Nguyen A., Lang D., Hoch J. A. Construction and properties of an integrable plasmid for Bacillus subtilis. J Bacteriol. 1983 Jun;154(3):1513–1515. doi: 10.1128/jb.154.3.1513-1515.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Geisler U., Schumann W. Isolation of stress mutants of Bacillus subtilis by a novel genetic method. FEMS Microbiol Lett. 1993 Apr 15;108(3):251–254. doi: 10.1111/j.1574-6968.1993.tb06110.x. [DOI] [PubMed] [Google Scholar]
  11. Gottesman S. Bacterial regulation: global regulatory networks. Annu Rev Genet. 1984;18:415–441. doi: 10.1146/annurev.ge.18.120184.002215. [DOI] [PubMed] [Google Scholar]
  12. Hanahan D. Studies on transformation of Escherichia coli with plasmids. J Mol Biol. 1983 Jun 5;166(4):557–580. doi: 10.1016/s0022-2836(83)80284-8. [DOI] [PubMed] [Google Scholar]
  13. Hecker M., Heim C., Völker U., Wölfel L. Induction of stress proteins by sodium chloride treatment in Bacillus subtilis. Arch Microbiol. 1988;150(6):564–566. doi: 10.1007/BF00408250. [DOI] [PubMed] [Google Scholar]
  14. Herman C., Ogura T., Tomoyasu T., Hiraga S., Akiyama Y., Ito K., Thomas R., D'Ari R., Bouloc P. Cell growth and lambda phage development controlled by the same essential Escherichia coli gene, ftsH/hflB. Proc Natl Acad Sci U S A. 1993 Nov 15;90(22):10861–10865. doi: 10.1073/pnas.90.22.10861. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Herman C., Thévenet D., D'Ari R., Bouloc P. Degradation of sigma 32, the heat shock regulator in Escherichia coli, is governed by HflB. Proc Natl Acad Sci U S A. 1995 Apr 11;92(8):3516–3520. doi: 10.1073/pnas.92.8.3516. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Jefferson R. A., Burgess S. M., Hirsh D. beta-Glucuronidase from Escherichia coli as a gene-fusion marker. Proc Natl Acad Sci U S A. 1986 Nov;83(22):8447–8451. doi: 10.1073/pnas.83.22.8447. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kreft J., Hughes C. Cloning vectors derived from plasmids and phage of Bacillus. Curr Top Microbiol Immunol. 1982;96:1–17. doi: 10.1007/978-3-642-68315-2_1. [DOI] [PubMed] [Google Scholar]
  18. Krüger E., Völker U., Hecker M. Stress induction of clpC in Bacillus subtilis and its involvement in stress tolerance. J Bacteriol. 1994 Jun;176(11):3360–3367. doi: 10.1128/jb.176.11.3360-3367.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Li M., Wong S. L. Cloning and characterization of the groESL operon from Bacillus subtilis. J Bacteriol. 1992 Jun;174(12):3981–3992. doi: 10.1128/jb.174.12.3981-3992.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Lindner C., Stülke J., Hecker M. Regulation of xylanolytic enzymes in Bacillus subtilis. Microbiology. 1994 Apr;140(Pt 4):753–757. doi: 10.1099/00221287-140-4-753. [DOI] [PubMed] [Google Scholar]
  21. Lucht J. M., Bremer E. Adaptation of Escherichia coli to high osmolarity environments: osmoregulation of the high-affinity glycine betaine transport system proU. FEMS Microbiol Rev. 1994 May;14(1):3–20. doi: 10.1111/j.1574-6976.1994.tb00067.x. [DOI] [PubMed] [Google Scholar]
  22. McCann M. P., Kidwell J. P., Matin A. The putative sigma factor KatF has a central role in development of starvation-mediated general resistance in Escherichia coli. J Bacteriol. 1991 Jul;173(13):4188–4194. doi: 10.1128/jb.173.13.4188-4194.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Measures J. C. Role of amino acids in osmoregulation of non-halophilic bacteria. Nature. 1975 Oct 2;257(5525):398–400. doi: 10.1038/257398a0. [DOI] [PubMed] [Google Scholar]
  24. Nilsson D., Lauridsen A. A., Tomoyasu T., Ogura T. A Lactococcus lactis gene encodes a membrane protein with putative ATPase activity that is homologous to the essential Escherichia coli ftsH gene product. Microbiology. 1994 Oct;140(Pt 10):2601–2610. doi: 10.1099/00221287-140-10-2601. [DOI] [PubMed] [Google Scholar]
  25. Ogasawara N., Nakai S., Yoshikawa H. Systematic sequencing of the 180 kilobase region of the Bacillus subtilis chromosome containing the replication origin. DNA Res. 1994;1(1):1–14. doi: 10.1093/dnares/1.1.1. [DOI] [PubMed] [Google Scholar]
  26. Rechsteiner M., Hoffman L., Dubiel W. The multicatalytic and 26 S proteases. J Biol Chem. 1993 Mar 25;268(9):6065–6068. [PubMed] [Google Scholar]
  27. Riethdorf S., Völker U., Gerth U., Winkler A., Engelmann S., Hecker M. Cloning, nucleotide sequence, and expression of the Bacillus subtilis lon gene. J Bacteriol. 1994 Nov;176(21):6518–6527. doi: 10.1128/jb.176.21.6518-6527.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Saito H., Shibata T., Ando T. Mapping of genes determining nonpermissiveness and host-specific restriction to bacteriophages in Bacillus subtilis Marburg. Mol Gen Genet. 1979 Feb 26;170(2):117–122. doi: 10.1007/BF00337785. [DOI] [PubMed] [Google Scholar]
  29. 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]
  30. Santos D., De Almeida D. F. Isolation and characterization of a new temperature-sensitive cell division mutant of Escherichia coli K-12. J Bacteriol. 1975 Dec;124(3):1502–1507. doi: 10.1128/jb.124.3.1502-1507.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Schulz A., Tzschaschel B., Schumann W. Isolation and analysis of mutants of the dnaK operon of Bacillus subtilis. Mol Microbiol. 1995 Feb;15(3):421–429. doi: 10.1111/j.1365-2958.1995.tb02256.x. [DOI] [PubMed] [Google Scholar]
  32. Tomoyasu T., Yamanaka K., Murata K., Suzaki T., Bouloc P., Kato A., Niki H., Hiraga S., Ogura T. Topology and subcellular localization of FtsH protein in Escherichia coli. J Bacteriol. 1993 Mar;175(5):1352–1357. doi: 10.1128/jb.175.5.1352-1357.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Tomoyasu T., Yuki T., Morimura S., Mori H., Yamanaka K., Niki H., Hiraga S., Ogura T. The Escherichia coli FtsH protein is a prokaryotic member of a protein family of putative ATPases involved in membrane functions, cell cycle control, and gene expression. J Bacteriol. 1993 Mar;175(5):1344–1351. doi: 10.1128/jb.175.5.1344-1351.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Völker U., Engelmann S., Maul B., Riethdorf S., Völker A., Schmid R., Mach H., Hecker M. Analysis of the induction of general stress proteins of Bacillus subtilis. Microbiology. 1994 Apr;140(Pt 4):741–752. doi: 10.1099/00221287-140-4-741. [DOI] [PubMed] [Google Scholar]
  35. Völker U., Mach H., Schmid R., Hecker M. Stress proteins and cross-protection by heat shock and salt stress in Bacillus subtilis. J Gen Microbiol. 1992 Oct;138(10):2125–2135. doi: 10.1099/00221287-138-10-2125. [DOI] [PubMed] [Google Scholar]
  36. Wetzstein M., Völker U., Dedio J., Löbau S., Zuber U., Schiesswohl M., Herget C., Hecker M., Schumann W. Cloning, sequencing, and molecular analysis of the dnaK locus from Bacillus subtilis. J Bacteriol. 1992 May;174(10):3300–3310. doi: 10.1128/jb.174.10.3300-3310.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Whatmore A. M., Chudek J. A., Reed R. H. The effects of osmotic upshock on the intracellular solute pools of Bacillus subtilis. J Gen Microbiol. 1990 Dec;136(12):2527–2535. doi: 10.1099/00221287-136-12-2527. [DOI] [PubMed] [Google Scholar]
  38. Wolfe K. H. Similarity between putative ATP-binding sites in land plant plastid ORF2280 proteins and the FtsH/CDC48 family of ATPases. Curr Genet. 1994 Apr;25(4):379–383. doi: 10.1007/BF00351493. [DOI] [PubMed] [Google Scholar]
  39. 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]
  40. Zuber U., Schumann W. Tn5cos: a transposon for restriction mapping of large plasmids using phage lambda terminase. Gene. 1991 Jul 15;103(1):69–72. doi: 10.1016/0378-1119(91)90392-o. [DOI] [PubMed] [Google Scholar]

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