Skip to main content
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1985 Nov;164(2):639–645. doi: 10.1128/jb.164.2.639-645.1985

Structure of two divergent promoters located in front of the gene encoding pullulanase in Klebsiella pneumoniae and positively regulated by the malT product.

C Chapon, O Raibaud
PMCID: PMC214300  PMID: 3902792

Abstract

Pullulanase is an extracellular starch-debranching enzyme produced by Klebsiella pneumoniae. When its structural gene, pulA, is introduced into Escherichia coli, it is controlled by malT, the positive regulator gene of the maltose regulon. Characterization of the region 5' to pulA and of the beginning of the gene described herein demonstrate that (i) pullulanase is probably a lipoprotein; (ii) an additional malT-controlled promoter (the malX promoter) lies adjacent to the pulA promoter and is oriented in the opposite direction; (iii) in common with the three previously described malT-controlled promoters, the pulA and malX promoters have a conserved hexanucleotide (consensus sequence, 5'-GGATGGA) 35 base pairs upstream from the transcription initiation site; and (iv) upstream from this conserved hexanucleotide the pulA and malX promoters differ from the other mal promoters in that they lack any detectable binding site for the cyclic AMP-binding protein.

Full text

PDF
639

Images in this article

Selected References

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

  1. Bedouelle H. Mutations in the promoter regions of the malEFG and malK-lamB operons of Escherichia coli K12. J Mol Biol. 1983 Nov 15;170(4):861–882. doi: 10.1016/s0022-2836(83)80192-2. [DOI] [PubMed] [Google Scholar]
  2. Bedouelle H., Schmeissner U., Hofnung M., Rosenberg M. Promoters of the malEFG and malK-lamB operons in Escherichia coli K12. J Mol Biol. 1982 Nov 15;161(4):519–531. doi: 10.1016/0022-2836(82)90405-3. [DOI] [PubMed] [Google Scholar]
  3. Casadaban M. J. Transposition and fusion of the lac genes to selected promoters in Escherichia coli using bacteriophage lambda and Mu. J Mol Biol. 1976 Jul 5;104(3):541–555. doi: 10.1016/0022-2836(76)90119-4. [DOI] [PubMed] [Google Scholar]
  4. Chapon C., Kolb A. Action of CAP on the malT promoter in vitro. J Bacteriol. 1983 Dec;156(3):1135–1143. doi: 10.1128/jb.156.3.1135-1143.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Clément J. M., Hofnung M. Gene sequence of the lambda receptor, an outer membrane protein of E. coli K12. Cell. 1981 Dec;27(3 Pt 2):507–514. doi: 10.1016/0092-8674(81)90392-5. [DOI] [PubMed] [Google Scholar]
  6. Debarbouille M., Cossart P., Raibaud O. A DNA sequence containing the control sites for gene malT and for the malPQ operon. Mol Gen Genet. 1982;185(1):88–92. doi: 10.1007/BF00333795. [DOI] [PubMed] [Google Scholar]
  7. Dessein A., Schwartz M. Is there a pullulanase in escherichia coli? Eur J Biochem. 1974 Jun 15;45(2):363–366. doi: 10.1111/j.1432-1033.1974.tb03561.x. [DOI] [PubMed] [Google Scholar]
  8. Duplay P., Bedouelle H., Fowler A., Zabin I., Saurin W., Hofnung M. Sequences of the malE gene and of its product, the maltose-binding protein of Escherichia coli K12. J Biol Chem. 1984 Aug 25;259(16):10606–10613. [PubMed] [Google Scholar]
  9. Débarbouillé M., Raibaud O. Expression of the Escherichia coli malPQ operon remains unaffected after drastic alteration of its promoter. J Bacteriol. 1983 Mar;153(3):1221–1227. doi: 10.1128/jb.153.3.1221-1227.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Débarbouillé M., Shuman H. A., Silhavy T. J., Schwartz M. Dominant constitutive mutations in malT, the positive regulator gene of the maltose regulon in Escherichia coli. J Mol Biol. 1978 Sep 15;124(2):359–371. doi: 10.1016/0022-2836(78)90304-2. [DOI] [PubMed] [Google Scholar]
  11. Eisele B., Rasched I. R., Wallenfels K. Molecular characterization of pullulanase from Aerobacter aerogenes. Eur J Biochem. 1972 Mar 15;26(1):62–67. doi: 10.1111/j.1432-1033.1972.tb01739.x. [DOI] [PubMed] [Google Scholar]
  12. Fried M., Crothers D. M. Equilibria and kinetics of lac repressor-operator interactions by polyacrylamide gel electrophoresis. Nucleic Acids Res. 1981 Dec 11;9(23):6505–6525. doi: 10.1093/nar/9.23.6505. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Froshauer S., Beckwith J. The nucleotide sequence of the gene for malF protein, an inner membrane component of the maltose transport system of Escherichia coli. Repeated DNA sequences are found in the malE-malF intercistronic region. J Biol Chem. 1984 Sep 10;259(17):10896–10903. [PubMed] [Google Scholar]
  14. Garner M. M., Revzin A. A gel electrophoresis method for quantifying the binding of proteins to specific DNA regions: application to components of the Escherichia coli lactose operon regulatory system. Nucleic Acids Res. 1981 Jul 10;9(13):3047–3060. doi: 10.1093/nar/9.13.3047. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Gilson E., Nikaido H., Hofnung M. Sequence of the malK gene in E.coli K12. Nucleic Acids Res. 1982 Nov 25;10(22):7449–7458. doi: 10.1093/nar/10.22.7449. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hope G. C., Dean A. C. Pullulanase synthesis in klebsiella (aerobacter) aerogenes strains growing in continuous culture. Biochem J. 1974 Nov;144(2):403–411. doi: 10.1042/bj1440403. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Michaelis S., Chapon C., D'Enfert C., Pugsley A. P., Schwartz M. Characterization and expression of the structural gene for pullulanase, a maltose-inducible secreted protein of Klebsiella pneumoniae. J Bacteriol. 1985 Nov;164(2):633–638. doi: 10.1128/jb.164.2.633-638.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Norrander J., Kempe T., Messing J. Construction of improved M13 vectors using oligodeoxynucleotide-directed mutagenesis. Gene. 1983 Dec;26(1):101–106. doi: 10.1016/0378-1119(83)90040-9. [DOI] [PubMed] [Google Scholar]
  19. Palm D., Goerl R., Burger K. J. Evolution of catalytic and regulatory sites in phosphorylases. Nature. 1985 Feb 7;313(6002):500–502. doi: 10.1038/313500a0. [DOI] [PubMed] [Google Scholar]
  20. Raibaud O., Débarbouillé M., Schwartz M. Use of deletions created in vitro to map transcriptional regulatory signals in the malA region of Escherichia coli. J Mol Biol. 1983 Jan 25;163(3):395–408. doi: 10.1016/0022-2836(83)90065-7. [DOI] [PubMed] [Google Scholar]
  21. Raibaud O., Gutierrez C., Schwartz M. Essential and nonessential sequences in malPp, a positively controlled promoter in Escherichia coli. J Bacteriol. 1985 Mar;161(3):1201–1208. doi: 10.1128/jb.161.3.1201-1208.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Raibaud O., Mock M., Schwartz M. A technique for integrating any DNA fragment into the chromosome of Escherichia coli. Gene. 1984 Jul-Aug;29(1-2):231–241. doi: 10.1016/0378-1119(84)90183-5. [DOI] [PubMed] [Google Scholar]
  23. Raibaud O., Schwartz M. Positive control of transcription initiation in bacteria. Annu Rev Genet. 1984;18:173–206. doi: 10.1146/annurev.ge.18.120184.001133. [DOI] [PubMed] [Google Scholar]
  24. Rosenberg M., Court D. Regulatory sequences involved in the promotion and termination of RNA transcription. Annu Rev Genet. 1979;13:319–353. doi: 10.1146/annurev.ge.13.120179.001535. [DOI] [PubMed] [Google Scholar]
  25. Rüther U., Koenen M., Otto K., Müller-Hill B. pUR222, a vector for cloning and rapid chemical sequencing of DNA. Nucleic Acids Res. 1981 Aug 25;9(16):4087–4098. doi: 10.1093/nar/9.16.4087. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Sanger F., Coulson A. R. The use of thin acrylamide gels for DNA sequencing. FEBS Lett. 1978 Mar 1;87(1):107–110. doi: 10.1016/0014-5793(78)80145-8. [DOI] [PubMed] [Google Scholar]
  27. 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]
  28. Takizawa N., Murooka Y. Cloning of the pullulanase gene and overproduction of pullulanase in Escherichia coli and Klebsiella aerogenes. Appl Environ Microbiol. 1985 Feb;49(2):294–298. doi: 10.1128/aem.49.2.294-298.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES