Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1984 Aug;159(2):658–662. doi: 10.1128/jb.159.2.658-662.1984

Genetic analysis of the functional relationship between colicin E3 and its immunity protein.

M Mock, C G Miyada, R J Collier
PMCID: PMC215694  PMID: 6086581

Abstract

Partial deletions in the immunity gene of the colicin E3 operon were used to study possible functions of the immunity protein besides protection against exogenous colicin. Nuclease BAL-31 was used to create a series of carboxyl-terminal deletions of the immunity gene. Mutants displaying lowered immunity against exogenous colicin were found, and six that had reduced but detectable levels of immunity were chosen for further analysis. DNA sequence analysis of the deletions showed that all six terminated within the last five codons of the immunity gene. The wild-type immunity gene was replaced by each of the six mutated immunity genes in a plasmid containing an otherwise functional colicin E3 operon. Transformants containing the resulting plasmids produced smaller colonies on solid medium and grew more slowly in liquid culture than transformants carrying the wild-type colicin and immunity genes. This result suggested that immunity protein was required to protect the cell against endogenous colicin E3. This idea was confirmed in experiments in which the colicin E3 and immunity genes were independently cloned on two compatible plasmid vectors.

Full text

PDF
659

Selected References

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

  1. 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]
  2. FREDERICQ P. Colicins and colicinogenic factors. Symp Soc Exp Biol. 1958;12:104–122. [PubMed] [Google Scholar]
  3. Helinski D. R., Herschman H. R. Effect of Rec mutations on the activity of colicinogenic factors. J Bacteriol. 1967 Sep;94(3):700–706. doi: 10.1128/jb.94.3.700-706.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Jakes K. S., Zinder N. D. Highly purified colicin E3 contains immunity protein. Proc Natl Acad Sci U S A. 1974 Sep;71(9):3380–3384. doi: 10.1073/pnas.71.9.3380. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Kretschmer P. J., Cohen S. N. Selected translocation of plasmid genes: frequency and regional specificity of translocation of the Tn3 element. J Bacteriol. 1977 May;130(2):888–899. doi: 10.1128/jb.130.2.888-899.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Little J. W., Mount D. W. The SOS regulatory system of Escherichia coli. Cell. 1982 May;29(1):11–22. doi: 10.1016/0092-8674(82)90085-x. [DOI] [PubMed] [Google Scholar]
  7. Masaki H., Ohta T. A plasmid region encoding the active fragment and the inhibitor protein of colicin E3--CA38. FEBS Lett. 1982 Nov 22;149(1):129–132. doi: 10.1016/0014-5793(82)81087-9. [DOI] [PubMed] [Google Scholar]
  8. Messing J., Vieira J. A new pair of M13 vectors for selecting either DNA strand of double-digest restriction fragments. Gene. 1982 Oct;19(3):269–276. doi: 10.1016/0378-1119(82)90016-6. [DOI] [PubMed] [Google Scholar]
  9. Mock M., Miyada C. G., Gunsalus R. P. Nucleotide sequence for the catalytic domain of colicin E3 and its immunity protein. Evidence for a third gene overlapping colicin. Nucleic Acids Res. 1983 Jun 11;11(11):3547–3557. doi: 10.1093/nar/11.11.3547. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Mock M., Pugsley A. P. The BtuB group col plasmids and homology between the colicins they encode. J Bacteriol. 1982 Jun;150(3):1069–1076. doi: 10.1128/jb.150.3.1069-1076.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Mock M., Schwartz M. Mutations which affect the structure and activity of colicin E3. J Bacteriol. 1980 May;142(2):384–390. doi: 10.1128/jb.142.2.384-390.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Ohno S., Ohno-Iwashita Y., Suzuki K., Imahori K. Purification and characterization of active component and active fragment of colicin E3. J Biochem. 1977 Oct;82(4):1045–1053. doi: 10.1093/oxfordjournals.jbchem.a131775. [DOI] [PubMed] [Google Scholar]
  13. Pugsley A. P., Schwartz M. A genetic approach to the study of mitomycin-induced lysis of Escherichia coli K-12 strains which produce colicin E2. Mol Gen Genet. 1983;190(3):366–372. doi: 10.1007/BF00331060. [DOI] [PubMed] [Google Scholar]
  14. 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]
  15. Schaller K., Nomura M. Colicin E2 is DNA endonuclease. Proc Natl Acad Sci U S A. 1976 Nov;73(11):3989–3993. doi: 10.1073/pnas.73.11.3989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. 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]
  17. Sutcliffe J. G. Complete nucleotide sequence of the Escherichia coli plasmid pBR322. Cold Spring Harb Symp Quant Biol. 1979;43(Pt 1):77–90. doi: 10.1101/sqb.1979.043.01.013. [DOI] [PubMed] [Google Scholar]
  18. 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]
  19. Watson R. J., Visentin L. P. Cloning of the ColE3-CA38 colicin and immunity genes and identification of a plasmid region which enhances colicin production. Gene. 1982 Sep;19(2):191–200. doi: 10.1016/0378-1119(82)90006-3. [DOI] [PubMed] [Google Scholar]

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

RESOURCES