Skip to main content
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1980 Nov;144(2):579–591. doi: 10.1128/jb.144.2.579-591.1980

Transcription of bacteriocinogenic plasmid CloDF13 in vivo and in vitro: structure of the cloacin immunity operon.

P J van den Elzen, R N Konings, E Veltkamp, H J Nijkamp
PMCID: PMC294705  PMID: 6159346

Abstract

Escherichia coli minicells harboring plasmid CloDF13 synthesized at least 25 messenger ribonucleic acid (RNA) species; three of these RNAs, a 2,400-, a 2,200-, and a 100-nucleotide RNA, were synthesized in relatively large amounts. Using insertion and deletion mutants of CloDF13 as well as an RNA blotting technique, we could demonstrate that these three RNAs are transcripts from the CloDF13 DNA region from 0 to 40%. This region contains the cloacin and immunity genes and the genetic information involved in plasmid DNA replication. A transcription map of this region is presented and discussed. The data indicate that the cloacin and immunity genes were coordinately transcribed into messenger RNAs of about 2,400 and 2,200 nucleotides, which differ in length at their 3' terminus. RNA polymerase binding studies and in vitro transcription assays indicated that transcription of these genes initiates at a promoter located around 32% on the CloDF13 map. Furthermore, it is shown that a 100-nucleotide RNA is encoded by the CloDF13 DNA region between 7.7 and 8.8% on the plasmid genome; the synthesis of this RNA proceeds in a direction opposite to the transcription of the cloacin and immunity genes.

Full text

PDF
579

Images in this article

Selected References

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

  1. Adler H. I., Fisher W. D., Cohen A., Hardigree A. A. MINIATURE escherichia coli CELLS DEFICIENT IN DNA. Proc Natl Acad Sci U S A. 1967 Feb;57(2):321–326. doi: 10.1073/pnas.57.2.321. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Alwine J. C., Kemp D. J., Stark G. R. Method for detection of specific RNAs in agarose gels by transfer to diazobenzyloxymethyl-paper and hybridization with DNA probes. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5350–5354. doi: 10.1073/pnas.74.12.5350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Andreoli P. M., Overbeeke N., Veltkamp E., van Embden J. D., Nijkamp H. J. Genetic map of the bacteriocinogenic plasmid CLO DF13 derived by insertion of the transposon Tn901. Mol Gen Genet. 1978 Mar 20;160(1):1–11. doi: 10.1007/BF00275113. [DOI] [PubMed] [Google Scholar]
  4. Bailey J. M., Davidson N. Methylmercury as a reversible denaturing agent for agarose gel electrophoresis. Anal Biochem. 1976 Jan;70(1):75–85. doi: 10.1016/s0003-2697(76)80049-8. [DOI] [PubMed] [Google Scholar]
  5. Calame K. L., Yamada Y., Shanblatt S. H., Nakada D. Location of promoter sites on plasmid NTP1 which contains the ampicillin resistance transposon Tn1701. J Mol Biol. 1979 Feb 5;127(4):397–409. doi: 10.1016/0022-2836(79)90229-8. [DOI] [PubMed] [Google Scholar]
  6. Clark A. J., Warren G. J. Conjugal transmission of plasmids. Annu Rev Genet. 1979;13:99–125. doi: 10.1146/annurev.ge.13.120179.000531. [DOI] [PubMed] [Google Scholar]
  7. De Wilde M., Davies J. E., Schmidt F. J. Low molecular weight RNA species encoded by a multiple drug resistance plasmid. Proc Natl Acad Sci U S A. 1978 Aug;75(8):3673–3677. doi: 10.1073/pnas.75.8.3673. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Edens L., van Wezenbeek P., Konings N. H., Schoenmakers J. G. Mapping of promoter sites on the genome of bacteriophage M13. Eur J Biochem. 1976 Nov 15;70(2):577–587. doi: 10.1111/j.1432-1033.1976.tb11049.x. [DOI] [PubMed] [Google Scholar]
  9. Koller T., Kübler O., Portmann R., Sogo J. M. High resolution physical mapping of specific binding sites of Escherichia coli RNA polymerase on the DNA of bacteriophage T7 . J Mol Biol. 1978 Mar 25;120(1):121–131. doi: 10.1016/0022-2836(78)90298-x. [DOI] [PubMed] [Google Scholar]
  10. Kolter R., Helinski D. R. Regulation of initiation of DNA replication. Annu Rev Genet. 1979;13:355–391. doi: 10.1146/annurev.ge.13.120179.002035. [DOI] [PubMed] [Google Scholar]
  11. Kool A. J., Nijkamp H. J. Isolation and characterization of a copy mutant of the bacteriocinogenic plasmid Clo DF13. J Bacteriol. 1974 Nov;120(2):569–578. doi: 10.1128/jb.120.2.569-578.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kool A. J., van Zeben M. S., Nijkamp H. J. Identification of messenger ribonucleic acids and proteins synthesized by the bacteriocinogenic factor Clo DF13 in purified minicells of Escherichia coli. J Bacteriol. 1974 Apr;118(1):213–224. doi: 10.1128/jb.118.1.213-224.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Laskey R. A., Mills A. D. Quantitative film detection of 3H and 14C in polyacrylamide gels by fluorography. Eur J Biochem. 1975 Aug 15;56(2):335–341. doi: 10.1111/j.1432-1033.1975.tb02238.x. [DOI] [PubMed] [Google Scholar]
  14. Maat J., Smith A. J. A method for sequencing restriction fragments with dideoxynucleoside triphosphates. Nucleic Acids Res. 1978 Dec;5(12):4537–4545. doi: 10.1093/nar/5.12.4537. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Maquat L. E., Reznikoff W. S. In vitro analysis of the Escherichia coli RNA polymerase interaction with wild-type and mutant lactose promoters. J Mol Biol. 1978 Nov 15;125(4):467–490. doi: 10.1016/0022-2836(78)90311-x. [DOI] [PubMed] [Google Scholar]
  16. Morita M., Oka A. The structure of a transcriptional unit on colicin E1 plasmid. Eur J Biochem. 1979 Jul;97(2):435–443. doi: 10.1111/j.1432-1033.1979.tb13131.x. [DOI] [PubMed] [Google Scholar]
  17. Oka A., Nomura N., Morita M., Sugisaki H., Sugimoto K., Takanami M. Nucleotide sequence of small ColE1 derivatives: structure of the regions essential for autonomous replication and colicin E1 immunity. Mol Gen Genet. 1979 May 4;172(2):151–159. doi: 10.1007/BF00268276. [DOI] [PubMed] [Google Scholar]
  18. Patient R. K. Characterization of in vitro transcription initiation and termination sites in Col E1 DNA. Nucleic Acids Res. 1979 Jun 25;6(8):2647–2665. doi: 10.1093/nar/6.8.2647. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. 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]
  20. Rivera M. J., Smits M. A., Quint W., Schoenmakers J. G., Konings R. N. Expression of bacteriophage M13 DNA in vivo. Localization of the transcription initiation and termination signal of the mRNA coding for the major capsid protein. Nucleic Acids Res. 1978 Aug;5(8):2895–2912. doi: 10.1093/nar/5.8.2895. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Smits M. A., Simons G., Konings R. N., Schoenmakers J. G. Expression of bacteriophage M13 dna in vivo. I. Synthesis of phage-specific RNA and protein in minicells. Biochim Biophys Acta. 1978 Nov 21;521(1):27–44. doi: 10.1016/0005-2787(78)90246-0. [DOI] [PubMed] [Google Scholar]
  22. Stuitje A. R., Veltkamp E., Maat J., Heyneker H. L. The nucleotide sequence surrounding the replication origin of the cop3 mutant of the bacteriocinogenic plasmid Clo DF13. Nucleic Acids Res. 1980 Apr 11;8(7):1459–1473. doi: 10.1093/nar/8.7.1459. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Stuitje A. R., Veltkamp E., Weijers P. J., Nijkamp H. J. Origin and direction of replication of the bacteriocinogenic plasmid Clo DF13. Nucleic Acids Res. 1979 Jan;6(1):71–80. doi: 10.1093/nar/6.1.71. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Stuitje A. R., Veltkamp E., van den Elzen P. J., Nijkamp H. J. In vitro construction of deletion mutants of the bacteriocinogenic plasmid Clo DF13. Nucleic Acids Res. 1978 Jun;5(6):1801–1820. doi: 10.1093/nar/5.6.1801. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Stüber D., Bujard H. Electron microscopy of DNA: determination of absolute molecular weights and linear density. Mol Gen Genet. 1977 Sep 9;154(3):299–303. doi: 10.1007/BF00571286. [DOI] [PubMed] [Google Scholar]
  26. Tabak H. F., Flavell R. A. A method for the recovery of DNA from agarose gels. Nucleic Acids Res. 1978 Jul;5(7):2321–2332. doi: 10.1093/nar/5.7.2321. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Tyler J., Sherratt D. J. Synthesis of E colicins in Escherichia coli. Mol Gen Genet. 1975 Oct 22;140(4):349–353. doi: 10.1007/BF00267325. [DOI] [PubMed] [Google Scholar]
  28. Van Ormondt H., Maat J., De Waard A., Van der Eb A. J. The nucleotide sequence of the transforming HpaI-E fragment of adenovirus type 5 DNA. Gene. 1978 Dec;4(4):309–328. doi: 10.1016/0378-1119(78)90048-3. [DOI] [PubMed] [Google Scholar]
  29. Veltkamp E., Barendsen W., Nijkamp H. J. Influence of protein and ribonucleic acid synthesis on the replication of the bacteriocinogenic factor Clo DF13 in Escherichia coli cells and minicells. J Bacteriol. 1974 Apr;118(1):165–174. doi: 10.1128/jb.118.1.165-174.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Veltkamp E., Nijkamp H. J. Characterization of a replication mutant of the bacteriocinogenic plasmid Clo DF13. Biochim Biophys Acta. 1976 Mar 17;425(3):356–367. doi: 10.1016/0005-2787(76)90263-x. [DOI] [PubMed] [Google Scholar]
  31. Veltkamp E., Nijkamp H. J. The role of DNA polymerase I, II and 3 in the replication of the bacteriocinogenic factor Clo DF 13. Mol Gen Genet. 1973 Sep 27;125(4):329–340. doi: 10.1007/BF00276588. [DOI] [PubMed] [Google Scholar]
  32. Wahl G. M., Stern M., Stark G. R. Efficient transfer of large DNA fragments from agarose gels to diazobenzyloxymethyl-paper and rapid hybridization by using dextran sulfate. Proc Natl Acad Sci U S A. 1979 Aug;76(8):3683–3687. doi: 10.1073/pnas.76.8.3683. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. van Embden J. D., Veltkamp E., Stuitje T., Andreoli P. M., Nijkamp H. J. Integration of a transposable DNA sequence which mediates ampicillin resistance into Clo DF13 plasmid DNA: determination of the site and orientation of TnA insertions. Plasmid. 1978 Feb;1(2):204–217. doi: 10.1016/0147-619x(78)90039-2. [DOI] [PubMed] [Google Scholar]
  34. van de Pol H., Veltkamp E., Nijkamp H. J. Clo DF13 plasmid genes affecting Flac transfer and propagation of male specific RNA phages. Mol Gen Genet. 1979 Jan 11;168(3):309–317. doi: 10.1007/BF00271501. [DOI] [PubMed] [Google Scholar]
  35. vd Pol H., Veltkamp E., Nijkamp H. J. Genetic information of the bacteriocinogenic plasmid Clo DF13 involved in the inhibition of the multiplication of double stranded DNA phages. Mol Gen Genet. 1980;178(3):535–540. doi: 10.1007/BF00337858. [DOI] [PubMed] [Google Scholar]
  36. von Gabain A., Bujard H. Interaction of Escherichia coli RNA polymerase with promoters of several coliphage and plasmid DNAs. Proc Natl Acad Sci U S A. 1979 Jan;76(1):189–193. doi: 10.1073/pnas.76.1.189. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES