Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1980 May;142(2):384–390. doi: 10.1128/jb.142.2.384-390.1980

Mutations which affect the structure and activity of colicin E3.

M Mock, M Schwartz
PMCID: PMC293982  PMID: 6155370

Abstract

Among 69 ColE3 mutant plasmids selected on the basis of their inability to produce an active colicin, seven (cop-1 to cop-7) were found to bear a mutation affecting the structural gene for colicin. Three of these (cop-1, cop-2 and cop-3) lead to the production of an inactive colicin molecule which has the same molecular weight as wild-type colicin E3 (67,000). These three inactive colicins are still able to interact with the outer membrane receptor. The cop-1 protein retained the ability to inhibit protein synthesis in vitro and therefore seems specifically affected in it ability to penetrate the cell envelope. The cop-2 and cop-3 proteins lost the ability to inhibit protein synthesis in vitro, and activity which is normally associated with the C-terminal part of the colicin molecule. On the basis of this and further evidence, it is suggested that the cop-2 and cop-3 mutations affect the structure of the C-terminal part of the colicin molecule. The other four mutations (cop-4 to cop-7) lead to the production of colicin-related polypeptides of lower molecular weight (29,000 to 45,000) which display a reaction of partial immunological identity with wild-type colicin. These four polypeptides are unable to interact with the cell surface receptor. Three of these mutants are shown to carry a nonsense mutation.

Full text

PDF
384

Images in this article

386Image
on p.386
387Image
on p.387
388Image
on p.388
388Image
on p.388

Selected References

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

  1. Andreoli P. M., Nijkamp H. J. Mutants of the Clo DF13 plasmid in Escherichia coli with a decreased bacteriocinogenic activity. Mol Gen Genet. 1976 Mar 22;144(2):159–170. doi: 10.1007/BF02428104. [DOI] [PubMed] [Google Scholar]
  2. Boon T. Inactivation of ribosomes in vitro by colicin E 3 . Proc Natl Acad Sci U S A. 1971 Oct;68(10):2421–2425. doi: 10.1073/pnas.68.10.2421. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bowman C. M., Dahlberg J. E., Ikemura T., Konisky J., Nomura M. Specific inactivation of 16S ribosomal RNA induced by colicin E3 in vivo. Proc Natl Acad Sci U S A. 1971 May;68(5):964–968. doi: 10.1073/pnas.68.5.964. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bowman C. M., Sidikaro J., Nomura M. Specific inactivation of ribosomes by colicin E3 in vitro and mechanism of immunity in colicinogenic cells. Nat New Biol. 1971 Dec 1;234(48):133–137. doi: 10.1038/newbio234133a0. [DOI] [PubMed] [Google Scholar]
  5. Gaastra W., Oudega B., de Graaf F. K. The use of mutants in the study of structure-function relationships in cloacin DF13. Biochim Biophys Acta. 1978 May 3;540(2):301–312. doi: 10.1016/0304-4165(78)90143-5. [DOI] [PubMed] [Google Scholar]
  6. Hardy K. G. Colicinogeny and related phenomena. Bacteriol Rev. 1975 Dec;39(4):464–515. doi: 10.1128/br.39.4.464-515.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Herschman H. R., Helinski D. R. Purification and characterization of colicin E2 and colicin E3. J Biol Chem. 1967 Nov 25;242(22):5360–5368. [PubMed] [Google Scholar]
  8. Hirose A., Kumagai J., Imahori K. Dessociation and reconstitution of colicin E3 and immunity substance complex. J Biochem. 1976 Feb;79(2):305–311. doi: 10.1093/oxfordjournals.jbchem.a131073. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  11. Lau C., Richards F. M. Proteolytic and chemical modification of colicin E3 activity. Biochemistry. 1976 Aug 24;15(17):3856–3863. doi: 10.1021/bi00662a032. [DOI] [PubMed] [Google Scholar]
  12. Mock M., Schwartz M. Mechanism of colicin E3 production in strains harboring wild-type or mutant plasmids. J Bacteriol. 1978 Nov;136(2):700–707. doi: 10.1128/jb.136.2.700-707.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. 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]
  14. Sabet S. F., Schnaitman C. A. Purification and properties of the colicin E3 receptor of Escherichia coli. J Biol Chem. 1973 Mar 10;248(5):1797–1806. [PubMed] [Google Scholar]
  15. Senior B. W., Holland I. B. Effect of colicin E3 upon the 30S ribosomal subunit of Escherichia coli. Proc Natl Acad Sci U S A. 1971 May;68(5):959–963. doi: 10.1073/pnas.68.5.959. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Tilby M., Hindennach I., Henning U. Bypass of receptor-mediated resistance to colicin E3 in Escherichia coli K-12. J Bacteriol. 1978 Dec;136(3):1189–1191. doi: 10.1128/jb.136.3.1189-1191.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. 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]
  18. Watson D. H., Sherratt D. J. In vivo proteolytic cleavage of colicins requires specific receptor binding. Nature. 1979 Mar 22;278(5702):362–364. doi: 10.1038/278362a0. [DOI] [PubMed] [Google Scholar]
  19. Webster R. E., Engelhardt D. L., Zinder N. D., Konigsberg W. Amber mutants and chain termination in vitro. J Mol Biol. 1967 Oct 14;29(1):27–43. doi: 10.1016/0022-2836(67)90179-9. [DOI] [PubMed] [Google Scholar]
  20. de Graaf F. K., Stukart M. J., Boogerd F. C., Metselaar K. Limited proteolysis of cloacin DF13 and characterization of the cleavage products. Biochemistry. 1978 Mar 21;17(6):1137–1142. doi: 10.1021/bi00599a031. [DOI] [PubMed] [Google Scholar]

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

RESOURCES