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. 1986 Jun;166(3):769–778. doi: 10.1128/jb.166.3.769-778.1986

Transcriptional analysis of beta-lactamase regulation in Bacillus licheniformis.

A J Salerno, J O Lampen
PMCID: PMC215193  PMID: 2423503

Abstract

The expression of the blaP gene for the beta-lactamase of Bacillus licheniformis was examined by transcriptional analyses. Radiolabeled probes containing the blaP gene or various regions 3' or 5' to it were used to analyze RNA samples prepared from induced and uninduced cultures of wild-type and mutant B. licheniformis strains. The level of blaP mRNA was low in uninduced wild-type cells. At 37 degrees C, blaP mRNA levels reached a maximum 1 h after induction while rising up to 180-fold and then declined, but remained severalfold above the uninduced level for several hours. The rate of beta-lactamase synthesis was roughly proportional to the levels of blaP mRNA in both wild-type and mutant strains, indicating that regulation of beta-lactamase formation occurs primarily at the level of transcription. Turnover of blaP mRNA in the presence of rifampin was rapid, giving a blaP mRNA half-life of about 2 min. Yet, high levels of blaP mRNA were maintained for at least 1 h after removal of free inducer. Three blaP mRNAs of 1.2, 2.9, and 3.4 kilobases were produced from the blaP promoter. The most abundant made up about 97% of all blaP transcripts and was also the smallest, ending at a transcriptional terminator located about 60 bases 3' to the blaP structural gene. Variables such as incubation temperature, cytotoxicity of inducer, and type of strain had essentially no effect on the ratio of large blaP mRNA to total blaP mRNA. The 2.9- and 3.4-kilobase blaP mRNAs identify potential locations of genetically linked regulators of beta-lactamase synthesis.

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Selected References

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

  1. Aiba H., Adhya S., de Crombrugghe B. Evidence for two functional gal promoters in intact Escherichia coli cells. J Biol Chem. 1981 Nov 25;256(22):11905–11910. [PubMed] [Google Scholar]
  2. Alwine J. C., Kemp D. J., Parker B. A., Reiser J., Renart J., Stark G. R., Wahl G. M. Detection of specific RNAs or specific fragments of DNA by fractionation in gels and transfer to diazobenzyloxymethyl paper. Methods Enzymol. 1979;68:220–242. doi: 10.1016/0076-6879(79)68017-5. [DOI] [PubMed] [Google Scholar]
  3. Ambler R. P. The structure of beta-lactamases. Philos Trans R Soc Lond B Biol Sci. 1980 May 16;289(1036):321–331. doi: 10.1098/rstb.1980.0049. [DOI] [PubMed] [Google Scholar]
  4. Aviv H., Leder P. Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid-cellulose. Proc Natl Acad Sci U S A. 1972 Jun;69(6):1408–1412. doi: 10.1073/pnas.69.6.1408. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Berk A. J., Sharp P. A. Sizing and mapping of early adenovirus mRNAs by gel electrophoresis of S1 endonuclease-digested hybrids. Cell. 1977 Nov;12(3):721–732. doi: 10.1016/0092-8674(77)90272-0. [DOI] [PubMed] [Google Scholar]
  6. Bettinger G. E., Lampen J. O. Penicillinase (beta-lactamase) induction in Bacillus licheniformis under Pseudogratuitous conditions by 2-(2'-carboxyphenyl)-benzoyl-6-aminopenicillanic acid. J Bacteriol. 1970 Oct;104(1):283–288. doi: 10.1128/jb.104.1.283-288.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. 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]
  9. DUERKSEN J. D. LOCALIZATION OF THE SITE OF FIXATION OF THE INDUCER, PENICILLIN, IN BACILLUS CEREUS. Biochim Biophys Acta. 1964 May 18;87:123–140. doi: 10.1016/0926-6550(64)90053-2. [DOI] [PubMed] [Google Scholar]
  10. Davies J. W. The stability and cell content of penicillinase messenger RNA in Bacillus licheniformis. J Gen Microbiol. 1969 Dec;59(2):171–184. doi: 10.1099/00221287-59-2-171. [DOI] [PubMed] [Google Scholar]
  11. Dubnau D. A., Pollock M. R. The genetics of Bacillus licheniformis penicillinase: a preliminary analysis from studies on mutation and inter-strain and intra-strain transformations. J Gen Microbiol. 1965 Oct;41(1):7–21. doi: 10.1099/00221287-41-1-7. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. Hussain I., Tsukagoshi N., Udaka S. Characterization of polyadenylated RNA in a protein-producing bacterium, Bacillus brevis 47. J Bacteriol. 1982 Sep;151(3):1162–1170. doi: 10.1128/jb.151.3.1162-1170.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. 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]
  15. 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]
  16. Jaurin B., Grundström T. ampC cephalosporinase of Escherichia coli K-12 has a different evolutionary origin from that of beta-lactamases of the penicillinase type. Proc Natl Acad Sci U S A. 1981 Aug;78(8):4897–4901. doi: 10.1073/pnas.78.8.4897. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. 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]
  18. 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]
  19. Koshland D., Sauer R. T., Botstein D. Diverse effects of mutations in the signal sequence on the secretion of beta-lactamase in Salmonella typhimurium. Cell. 1982 Oct;30(3):903–914. doi: 10.1016/0092-8674(82)90295-1. [DOI] [PubMed] [Google Scholar]
  20. Kroyer J., Chang S. The promoter-proximal region of the Bacillus licheniformis penicillinase gene: Nucleotide sequence and predicted leader peptide sequence. Gene. 1981 Dec;15(4):343–347. doi: 10.1016/0378-1119(81)90177-3. [DOI] [PubMed] [Google Scholar]
  21. 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]
  22. Maniatis T., Jeffrey A., Kleid D. G. Nucleotide sequence of the rightward operator of phage lambda. Proc Natl Acad Sci U S A. 1975 Mar;72(3):1184–1188. doi: 10.1073/pnas.72.3.1184. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Maniatis T., Jeffrey A., van deSande H. Chain length determination of small double- and single-stranded DNA molecules by polyacrylamide gel electrophoresis. Biochemistry. 1975 Aug 26;14(17):3787–3794. doi: 10.1021/bi00688a010. [DOI] [PubMed] [Google Scholar]
  24. 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]
  25. 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]
  26. McMaster G. K., Carmichael G. G. Analysis of single- and double-stranded nucleic acids on polyacrylamide and agarose gels by using glyoxal and acridine orange. Proc Natl Acad Sci U S A. 1977 Nov;74(11):4835–4838. doi: 10.1073/pnas.74.11.4835. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Mézes P. S., Wang W., Yeh E. C., Lampen J. O. Construction of penP delta 1, Bacillus licheniformis 749/C beta-lactamase lacking site for lipoprotein modification. Expression in Escherichia coli and Bacillus subtilis. J Biol Chem. 1983 Sep 25;258(18):11211–11218. [PubMed] [Google Scholar]
  28. 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]
  29. POLLOCK M. R., PERRET C. J. The relation between fixation of penicillin sulphur and penicillinase adaptation in B cereus. Br J Exp Pathol. 1951 Oct;32(5):387–396. [PMC free article] [PubMed] [Google Scholar]
  30. Radloff R., Bauer W., Vinograd J. A dye-buoyant-density method for the detection and isolation of closed circular duplex DNA: the closed circular DNA in HeLa cells. Proc Natl Acad Sci U S A. 1967 May;57(5):1514–1521. doi: 10.1073/pnas.57.5.1514. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  32. Saino Y., Kobayashi F., Inoue M., Mitsuhashi S. Purification and properties of inducible penicillin beta-lactamase isolated from Pseudomonas maltophilia. Antimicrob Agents Chemother. 1982 Oct;22(4):564–570. doi: 10.1128/aac.22.4.564. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Sargent M. G. Rapid fixed-time assay for penicillinase. J Bacteriol. 1968 Apr;95(4):1493–1494. doi: 10.1128/jb.95.4.1493-1494.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. 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]
  35. Thomas P. S. Hybridization of denatured RNA transferred or dotted nitrocellulose paper. Methods Enzymol. 1983;100:255–266. doi: 10.1016/0076-6879(83)00060-9. [DOI] [PubMed] [Google Scholar]
  36. Vieira J., Messing J. The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene. 1982 Oct;19(3):259–268. doi: 10.1016/0378-1119(82)90015-4. [DOI] [PubMed] [Google Scholar]
  37. Weaver R. F., Weissmann C. Mapping of RNA by a modification of the Berk-Sharp procedure: the 5' termini of 15 S beta-globin mRNA precursor and mature 10 s beta-globin mRNA have identical map coordinates. Nucleic Acids Res. 1979 Nov 10;7(5):1175–1193. doi: 10.1093/nar/7.5.1175. [DOI] [PMC free article] [PubMed] [Google Scholar]

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