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. 1986 Dec;168(3):1128–1132. doi: 10.1128/jb.168.3.1128-1132.1986

Nucleotide sequence of the penicillinase repressor gene penI of Bacillus licheniformis and regulation of penP and penI by the repressor.

T Himeno, T Imanaka, S Aiba
PMCID: PMC213612  PMID: 3096969

Abstract

Bacillus licheniformis penicillinase genes, penP and penI, are coded on a 4.2-kilobase EcoRI fragment of pTTE21 (T. Imanaka, T. Tanaka, H. Tsunekawa, and S. Aiba, J. Bacteriol. 147:776-186, 1981). The EcoRI fragment was subcloned in a low-copy-number plasmid pTB522 in Bacillus subtilis. B. subtilis carrying the recombinant plasmid pPTB60 (Tcr penP+ penI+) was chemically mutagenized. Of about 150,000 colonies, two penI(Ts) mutant plasmids, pPTB60D13 and pPTB60E24, were screened by the plate assay at 30 and 48 degrees C for penicillinase. By constructing recombinant plasmids between wild-type and mutant plasmids, the mutation points were shown to be located in a 1.7-kilobase EcoRI-PstI fragment. The EcoRI-PstI fragments of the wild-type plasmid and two mutant plasmids were sequenced. A large open reading frame, composed of 384 bases and 128 amino acid residues (molecular weight, 14,983), was found. Since the mutation points were located at different positions in the protein coding region (Ala to Val for pPTB60D13 and Pro to Leu for pPTB60E24), the coding region was concluded to be the penI gene. A Shine-Dalgarno sequence was found 7 bases upstream from the translation start site (ATG). A probable promoter sequence which is very similar to the consensus sequence was also found upstream of the penP promoter, but in the opposite direction. A consensus twofold symmetric sequence (AAAGTATTA CATATGTAAGNTTT) which might have been used as a repressor binding region was found downstream and in the midst of the penP promoter and also downstream of the penI promoter. The regulation of penP and penI by the repressor is discussed.

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Selected References

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  1. Brammar W. J., Muir S., McMorris A. Molecular cloning of the gene for the beta-lactamase of Bacillus licheniformis and its expression in Escherichia coli. Mol Gen Genet. 1980 Apr;178(1):217–224. doi: 10.1007/BF00267232. [DOI] [PubMed] [Google Scholar]
  2. Brent R., Ptashne M. Mechanism of action of the lexA gene product. Proc Natl Acad Sci U S A. 1981 Jul;78(7):4204–4208. doi: 10.1073/pnas.78.7.4204. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. DiLauro R., Taniguchi T., Musso R., de Crombrugghe B. Unusual location and function of the operator in the Escherichia coli galactose operon. Nature. 1979 Jun 7;279(5713):494–500. doi: 10.1038/279494a0. [DOI] [PubMed] [Google Scholar]
  4. Dunaway M., Manly S. P., Matthews K. S. Model for lactose repressor protein and its interaction with ligands. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7181–7185. doi: 10.1073/pnas.77.12.7181. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Gray O., Chang S. Molecular cloning and expression of Bacillus licheniformis beta-lactamase gene in Escherichia coli and Bacillus subtilis. J Bacteriol. 1981 Jan;145(1):422–428. doi: 10.1128/jb.145.1.422-428.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Gunsalus R. P., Yanofsky C. Nucleotide sequence and expression of Escherichia coli trpR, the structural gene for the trp aporepressor. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7117–7121. doi: 10.1073/pnas.77.12.7117. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Imanaka T., Fujii M., Aiba S. Isolation and characterization of antibiotic resistance plasmids from thermophilic bacilli and construction of deletion plasmids. J Bacteriol. 1981 Jun;146(3):1091–1097. doi: 10.1128/jb.146.3.1091-1097.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Imanaka T., Fujii M., Aramori I., Aiba S. Transformation of Bacillus stearothermophilus with plasmid DNA and characterization of shuttle vector plasmids between Bacillus stearothermophilus and Bacillus subtilis. J Bacteriol. 1982 Mar;149(3):824–830. doi: 10.1128/jb.149.3.824-830.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Imanaka T., Himeno T., Aiba S. Effect of in vitro DNA rearrangement in the NH2-terminal region of the penicillinase gene from Bacillus licheniformis on the mode of expression in Bacillus subtilis. J Gen Microbiol. 1985 Jul;131(7):1753–1763. doi: 10.1099/00221287-131-7-1753. [DOI] [PubMed] [Google Scholar]
  10. Imanaka T., Tanaka T., Tsunekawa H., Aiba S. Cloning of the genes for penicillinase, penP and penI, of Bacillus licheniformis in some vector plasmids and their expression in Escherichia coli, Bacillus subtilis, and Bacillus licheniformis. J Bacteriol. 1981 Sep;147(3):776–786. doi: 10.1128/jb.147.3.776-786.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Imsande J. Genetic regulation of penicillinase synthesis in Gram-positive bacteria. Microbiol Rev. 1978 Mar;42(1):67–83. doi: 10.1128/mr.42.1.67-83.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Isackson P. J., Bertrand K. P. Dominant negative mutations in the Tn10 tet repressor: evidence for use of the conserved helix-turn-helix motif in DNA binding. Proc Natl Acad Sci U S A. 1985 Sep;82(18):6226–6230. doi: 10.1073/pnas.82.18.6226. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kelly L. E., Brammar W. J. A frameshift mutation that elongates the penicillinase protein of Bacillus licheniformis. J Mol Biol. 1973 Oct 15;80(1):135–147. doi: 10.1016/0022-2836(73)90237-4. [DOI] [PubMed] [Google Scholar]
  14. Kelly L. E., Brammar W. J. The polycistronic nature of the penicillinase structural and regulatory genes in Bacillus licheniformis. J Mol Biol. 1973 Oct 15;80(1):149–154. doi: 10.1016/0022-2836(73)90238-6. [DOI] [PubMed] [Google Scholar]
  15. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  16. McLaughlin J. R., Chang S. Y., Chang S. Transcriptional analyses of the Bacillus licheniformis penP gene. Nucleic Acids Res. 1982 Jul 10;10(13):3905–3919. doi: 10.1093/nar/10.13.3905. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. McLaughlin J. R., Murray C. L., Rabinowitz J. C. Unique features in the ribosome binding site sequence of the gram-positive Staphylococcus aureus beta-lactamase gene. J Biol Chem. 1981 Nov 10;256(21):11283–11291. [PubMed] [Google Scholar]
  18. Messing J. New M13 vectors for cloning. Methods Enzymol. 1983;101:20–78. doi: 10.1016/0076-6879(83)01005-8. [DOI] [PubMed] [Google Scholar]
  19. Moran C. P., Jr, Lang N., LeGrice S. F., Lee G., Stephens M., Sonenshein A. L., Pero J., Losick R. Nucleotide sequences that signal the initiation of transcription and translation in Bacillus subtilis. Mol Gen Genet. 1982;186(3):339–346. doi: 10.1007/BF00729452. [DOI] [PubMed] [Google Scholar]
  20. Neugebauer K., Sprengel R., Schaller H. Penicillinase from Bacillus licheniformis: nucleotide sequence of the gene and implications for the biosynthesis of a secretory protein in a Gram-positive bacterium. Nucleic Acids Res. 1981 Jun 11;9(11):2577–2588. doi: 10.1093/nar/9.11.2577. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Sherratt D. J., Collins J. F. Analysis by transformation of the penicillinase system in Bacillus licheniformis. J Gen Microbiol. 1973 May;76(1):217–230. doi: 10.1099/00221287-76-1-217. [DOI] [PubMed] [Google Scholar]
  22. Wertman K. F., Mount D. W. Nucleotide sequence binding specificity of the LexA repressor of Escherichia coli K-12. J Bacteriol. 1985 Jul;163(1):376–384. doi: 10.1128/jb.163.1.376-384.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. de Crombrugghe B., Busby S., Buc H. Cyclic AMP receptor protein: role in transcription activation. Science. 1984 May 25;224(4651):831–838. doi: 10.1126/science.6372090. [DOI] [PubMed] [Google Scholar]

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