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. 1985 Dec;82(24):8389–8393. doi: 10.1073/pnas.82.24.8389

Structural and functional organization of the colicin E1 operon.

N S Waleh, P H Johnson
PMCID: PMC390921  PMID: 3936034

Abstract

We analyzed the structural and functional relationships among independently cloned segments of the plasmid ColE1 region that regulates and codes for colicin E1 (cea), immunity (imm), and the mitomycin C-induced lethality function (lys). On the basis of physiological properties, restriction endonuclease mapping, and DNA sequence analysis, the following recombinant plasmids were determined to represent three major functional classes: pNP12 (cea+, imm+, lys+), pNP4 (cea+, imm+, lys-), and pNP6 (cea+, imm-, lys-). Our results have established the order, boundaries, and relative orientation of the three structural genes, the location of the promoter region for imm gene transcription, and the predicted amino acid sequences of the imm and lys gene products. Hydropathicity analysis suggests that both proteins have hydrophobic amino acid segments characteristic of membrane-associated proteins. A model for the structure and expression of the colicin E1 operon is proposed in which the cea and lys genes are expressed from a single inducible promoter that is controlled by the lexA repressor in response to the SOS system of Escherichia coli. The imm gene lies between the cea and lys genes and is expressed by transcription in the opposite direction from a promoter located within the lys gene. This arrangement of structural genes indicates that the transcriptional units for all three genes overlap. We suggest that the formation of anti-sense RNA may be an important element in the coordinate regulation of gene expression in this system.

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Selected References

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  1. Bochner B. R., Huang H. C., Schieven G. L., Ames B. N. Positive selection for loss of tetracycline resistance. J Bacteriol. 1980 Aug;143(2):926–933. doi: 10.1128/jb.143.2.926-933.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Chen H. Z., Zubay G. Analysis of ColE1 expression in vitro after chromosome fragmentation. J Bacteriol. 1983 May;154(2):650–655. doi: 10.1128/jb.154.2.650-655.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Davidson V. L., Cramer W. A., Bishop L. J., Brunden K. R. Dependence of the activity of colicin E1 in artificial membrane vesicles on pH, membrane potential, and vesicle size. J Biol Chem. 1984 Jan 10;259(1):594–600. [PubMed] [Google Scholar]
  4. Davies J. K., Reeves P. Genetics of resistance to colicins in Escherichia coli K-12: cross-resistance among colicins of group A. J Bacteriol. 1975 Jul;123(1):102–117. doi: 10.1128/jb.123.1.102-117.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Davies J. K., Reeves P. Genetics of resistance to colicins in Escherichia coli K-12: cross-resistance among colicins of group B. J Bacteriol. 1975 Jul;123(1):96–101. doi: 10.1128/jb.123.1.96-101.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Di Masi D. R., White J. C., Schnaitman C. A., Bradbeer C. Transport of vitamin B12 in Escherichia coli: common receptor sites for vitamin B12 and the E colicins on the outer membrane of the cell envelope. J Bacteriol. 1973 Aug;115(2):506–513. doi: 10.1128/jb.115.2.506-513.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dougan G., Sherratt D. The transposon Tn1 as a probe for studying ColE1 structure and function. Mol Gen Genet. 1977 Mar 7;151(2):151–160. doi: 10.1007/BF00338689. [DOI] [PubMed] [Google Scholar]
  8. Ebina Y., Kishi F., Miki T., Kagamiyama H., Nakazawa T., Nakazawa A. The nucleotide sequence surrounding the promoter region of colicin E1 gene. Gene. 1981 Nov;15(2-3):119–126. doi: 10.1016/0378-1119(81)90121-9. [DOI] [PubMed] [Google Scholar]
  9. Ebina Y., Kishi F., Nakazawa A. Direct participation of lexA protein in repression of colicin E1 synthesis. J Bacteriol. 1982 Jun;150(3):1479–1481. doi: 10.1128/jb.150.3.1479-1481.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Ebina Y., Nakazawa A. Cyclic AMP-dependent initiation and rho-dependent termination of colicin E1 gene transcription. J Biol Chem. 1983 Jun 10;258(11):7072–7078. [PubMed] [Google Scholar]
  11. Ebina Y., Takahara Y., Kishi F., Nakazawa A., Brent R. LexA protein is a repressor of the colicin E1 gene. J Biol Chem. 1983 Nov 10;258(21):13258–13261. [PubMed] [Google Scholar]
  12. Hakkaart M. J., Veltkamp E., Nijkamp H. J. Protein H encoded by plasmid Clo DF13 involved in lysis of the bacterial host. I. Localisation of the gene and identification and subcellular localisation of the gene H product. Mol Gen Genet. 1981;183(2):318–325. doi: 10.1007/BF00270635. [DOI] [PubMed] [Google Scholar]
  13. Holt G. R., Davis W. E., Ailor E. I., Warren A. H., Elyassi H. Massive airway hemorrhage after transtracheal aspiration. South Med J. 1978 Mar;71(3):325–327. doi: 10.1097/00007611-197803000-00031. [DOI] [PubMed] [Google Scholar]
  14. Inselburg J. Isolation, mapping, and examination of effects of TnA insertions in ColE1 plasmids. J Bacteriol. 1977 Jan;129(1):482–491. doi: 10.1128/jb.129.1.482-491.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kelly S. V., Johnson P. H. The relaxation complex of colicinogenic factor E1: electrophoretic properties and a novel assay system for sodium dodecyl sulfate-mediated induction. Anal Biochem. 1980 Sep 15;107(2):362–368. doi: 10.1016/0003-2697(80)90397-8. [DOI] [PubMed] [Google Scholar]
  16. Kyte J., Doolittle R. F. A simple method for displaying the hydropathic character of a protein. J Mol Biol. 1982 May 5;157(1):105–132. doi: 10.1016/0022-2836(82)90515-0. [DOI] [PubMed] [Google Scholar]
  17. Males B. M., Stocker B. A. Colicins E4, E5, E6 and A and properties of btuB+ colicinogenic transconjugants. J Gen Microbiol. 1982 Jan;128(1):95–106. doi: 10.1099/00221287-128-1-95. [DOI] [PubMed] [Google Scholar]
  18. OZEKI H., STOCKER B. A., DE MARGERIE H. Production of colicine by single bacteria. Nature. 1959 Aug 1;184:337–339. doi: 10.1038/184337a0. [DOI] [PubMed] [Google Scholar]
  19. OZEKI H., STOCKER B. A., SMITH S. M. Transmission of colicinogeny between strains of Salmonella typhimurium grown together. J Gen Microbiol. 1962 Sep;28:671–687. doi: 10.1099/00221287-28-4-671. [DOI] [PubMed] [Google Scholar]
  20. 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]
  21. 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]
  22. Schwartz S. A., Helinski D. R. Purification and characterization of colicin E1. J Biol Chem. 1971 Oct 25;246(20):6318–6327. [PubMed] [Google Scholar]
  23. Shafferman A., Flashner Y., Cohen S. ColE1 DNA sequences interacting in cis, essential for mitomycin-C induced lethality. Mol Gen Genet. 1979 Oct 2;176(1):139–146. doi: 10.1007/BF00334305. [DOI] [PubMed] [Google Scholar]
  24. Suit J. L., Fan M. L., Sabik J. F., Labarre R., Luria S. E. Alternative forms of lethality in mitomycin C-induced bacteria carrying ColE1 plasmids. Proc Natl Acad Sci U S A. 1983 Jan;80(2):579–583. doi: 10.1073/pnas.80.2.579. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Tessman E. S., Peterson P. K. tif-dependent induction of colicin E1, prophage lambda, and filamentation in Escherichia coli K-12. J Bacteriol. 1980 Sep;143(3):1307–1317. doi: 10.1128/jb.143.3.1307-1317.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Villa-Komaroff L., Efstratiadis A., Broome S., Lomedico P., Tizard R., Naber S. P., Chick W. L., Gilbert W. A bacterial clone synthesizing proinsulin. Proc Natl Acad Sci U S A. 1978 Aug;75(8):3727–3731. doi: 10.1073/pnas.75.8.3727. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Yamada M., Ebina Y., Miyata T., Nakazawa T., Nakazawa A. Nucleotide sequence of the structural gene for colicin E1 and predicted structure of the protein. Proc Natl Acad Sci U S A. 1982 May;79(9):2827–2831. doi: 10.1073/pnas.79.9.2827. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Yamada M., Nakazawa A. Factors necessary for the export process of colicin E1 across cytoplasmic membrane of Escherichia coli. Eur J Biochem. 1984 Apr 16;140(2):249–255. doi: 10.1111/j.1432-1033.1984.tb08095.x. [DOI] [PubMed] [Google Scholar]

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