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. 1993 Jun;175(11):3394–3400. doi: 10.1128/jb.175.11.3394-3400.1993

Sequence and molecular characterization of a DNA region encoding a small heat shock protein of Clostridium acetobutylicum.

U Sauer 1, P Dürre 1
PMCID: PMC204737  PMID: 8501044

Abstract

A DNA region of Clostridium acetobutylicum containing a gene (hsp 18) with significant homology to a family of small eukaryotic heat shock proteins was cloned and sequenced. It is the second reported sequence of a low-molecular-weight heat shock protein from gram-positive bacteria and is induced not only by heat shock but also at the onset of solventogenesis, as determined by Northern (RNA) blot analysis, thus confirming the results of an earlier study performed at the protein level (A. Pich, F. Narberhaus, and H. Bahl, Appl. Microbiol. Biotechnol. 33:697-704, 1990). By primer extension analysis, a transcriptional start site was identified 149 bp upstream of hsp18. This site was preceded by a region that exhibits high homology to the consensus promoter sequences of gram-positive bacteria, as well as sigma 70-dependent Escherichia coli. A direct repeat structure was detected in the -35 region. The promoter is located 196 bp from the start of a potential regulatory tRNA(Thr)ACG gene involved in the shift to solventogenesis which is transcribed in the opposite direction. A putative rho-independent transcription termination structure was identified at the 3' end of hsp18.

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Selected References

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  1. Allen S. P., Polazzi J. O., Gierse J. K., Easton A. M. Two novel heat shock genes encoding proteins produced in response to heterologous protein expression in Escherichia coli. J Bacteriol. 1992 Nov;174(21):6938–6947. doi: 10.1128/jb.174.21.6938-6947.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Booth R. J., Harris D. P., Love J. M., Watson J. D. Antigenic proteins of Mycobacterium leprae. Complete sequence of the gene for the 18-kDa protein. J Immunol. 1988 Jan 15;140(2):597–601. [PubMed] [Google Scholar]
  3. 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]
  4. Gerischer U., Dürre P. mRNA analysis of the adc gene region of Clostridium acetobutylicum during the shift to solventogenesis. J Bacteriol. 1992 Jan;174(2):426–433. doi: 10.1128/jb.174.2.426-433.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Gralla J. D. Transcriptional control--lessons from an E. coli promoter data base. Cell. 1991 Aug 9;66(3):415–418. doi: 10.1016/0092-8674(81)90001-5. [DOI] [PubMed] [Google Scholar]
  6. Graves M. C., Rabinowitz J. C. In vivo and in vitro transcription of the Clostridium pasteurianum ferredoxin gene. Evidence for "extended" promoter elements in gram-positive organisms. J Biol Chem. 1986 Aug 25;261(24):11409–11415. [PubMed] [Google Scholar]
  7. Jones D. T., Woods D. R. Acetone-butanol fermentation revisited. Microbiol Rev. 1986 Dec;50(4):484–524. doi: 10.1128/mr.50.4.484-524.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Killeen K. P., Nelson D. R. Acceleration of starvation- and glycerol-induced myxospore formation by prior heat shock in Myxococcus xanthus. J Bacteriol. 1988 Nov;170(11):5200–5207. doi: 10.1128/jb.170.11.5200-5207.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Kurtz S., Rossi J., Petko L., Lindquist S. An ancient developmental induction: heat-shock proteins induced in sporulation and oogenesis. Science. 1986 Mar 7;231(4742):1154–1157. doi: 10.1126/science.3511530. [DOI] [PubMed] [Google Scholar]
  10. Maniak M., Nellen W. A developmentally regulated membrane protein gene in Dictyostelium discoideum is also induced by heat shock and cold shock. Mol Cell Biol. 1988 Jan;8(1):153–159. doi: 10.1128/mcb.8.1.153. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Marck C. 'DNA Strider': a 'C' program for the fast analysis of DNA and protein sequences on the Apple Macintosh family of computers. Nucleic Acids Res. 1988 Mar 11;16(5):1829–1836. doi: 10.1093/nar/16.5.1829. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Mazodier P., Guglielmi G., Davies J., Thompson C. J. Characterization of the groEL-like genes in Streptomyces albus. J Bacteriol. 1991 Nov;173(22):7382–7386. doi: 10.1128/jb.173.22.7382-7386.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Nagao R. T., Czarnecka E., Gurley W. B., Schöffl F., Key J. L. Genes for low-molecular-weight heat shock proteins of soybeans: sequence analysis of a multigene family. Mol Cell Biol. 1985 Dec;5(12):3417–3428. doi: 10.1128/mcb.5.12.3417. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Narberhaus F., Bahl H. Cloning, sequencing, and molecular analysis of the groESL operon of Clostridium acetobutylicum. J Bacteriol. 1992 May;174(10):3282–3289. doi: 10.1128/jb.174.10.3282-3289.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Narberhaus F., Giebeler K., Bahl H. Molecular characterization of the dnaK gene region of Clostridium acetobutylicum, including grpE, dnaJ, and a new heat shock gene. J Bacteriol. 1992 May;174(10):3290–3299. doi: 10.1128/jb.174.10.3290-3299.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Nerland A. H., Mustafa A. S., Sweetser D., Godal T., Young R. A. A protein antigen of Mycobacterium leprae is related to a family of small heat shock proteins. J Bacteriol. 1988 Dec;170(12):5919–5921. doi: 10.1128/jb.170.12.5919-5921.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Platt T. Transcription termination and the regulation of gene expression. Annu Rev Biochem. 1986;55:339–372. doi: 10.1146/annurev.bi.55.070186.002011. [DOI] [PubMed] [Google Scholar]
  18. 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]
  19. Sauer U., Dürre P. Possible function of tRNA(Thr)ACG in regulation of solvent formation in Clostridium acetobutylicum. FEMS Microbiol Lett. 1992 Dec 15;100(1-3):147–153. doi: 10.1111/j.1574-6968.1992.tb14033.x. [DOI] [PubMed] [Google Scholar]
  20. Shine J., Dalgarno L. The 3'-terminal sequence of Escherichia coli 16S ribosomal RNA: complementarity to nonsense triplets and ribosome binding sites. Proc Natl Acad Sci U S A. 1974 Apr;71(4):1342–1346. doi: 10.1073/pnas.71.4.1342. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Terracciano J. S., Rapaport E., Kashket E. R. Stress- and Growth Phase-Associated Proteins of Clostridium acetobutylicum. Appl Environ Microbiol. 1988 Aug;54(8):1989–1995. doi: 10.1128/aem.54.8.1989-1995.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Tinoco I., Jr, Borer P. N., Dengler B., Levin M. D., Uhlenbeck O. C., Crothers D. M., Bralla J. Improved estimation of secondary structure in ribonucleic acids. Nat New Biol. 1973 Nov 14;246(150):40–41. doi: 10.1038/newbio246040a0. [DOI] [PubMed] [Google Scholar]
  23. Verbon A., Hartskeerl R. A., Schuitema A., Kolk A. H., Young D. B., Lathigra R. The 14,000-molecular-weight antigen of Mycobacterium tuberculosis is related to the alpha-crystallin family of low-molecular-weight heat shock proteins. J Bacteriol. 1992 Feb;174(4):1352–1359. doi: 10.1128/jb.174.4.1352-1359.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Walter K. A., Bennett G. N., Papoutsakis E. T. Molecular characterization of two Clostridium acetobutylicum ATCC 824 butanol dehydrogenase isozyme genes. J Bacteriol. 1992 Nov;174(22):7149–7158. doi: 10.1128/jb.174.22.7149-7158.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. 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]
  26. Young M., Minton N. P., Staudenbauer W. L. Recent advances in the genetics of the clostridia. FEMS Microbiol Rev. 1989 Dec;5(4):301–325. doi: 10.1111/j.1574-6968.1989.tb03402.x. [DOI] [PubMed] [Google Scholar]

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