Abstract
The mechanism of action of colicin E3 (E3) was investigated in an in vitro system. Purified ribosomes are less susceptible to E3 than crude or washed ribosomes. A factor was found in the supernatant fraction of normal Escherichia coli cells that stimulates inactivation of ribosomes by E3, and on addition of this factor, about one tenth as much E3 was required for inactivation of ribosomes. On heating a mixture of E3 and this factor above 60°, the ribosome inactivating activity of E3 increased greatly, and an amount corresponding to 0.01 μg of E3 was sufficient to inactivate 1.0 A260 unit of ribosomes completely. By this treatment bacteriocidal activity of E3 decreased considerably, as the ratio of the two activities of E3 (ribosome inactivating activity and bacteriocidal activity) increased to 6 × 104-fold. It is evident that the two activities do not run in parallel.
This heat-treated product cleaved 16S rRNA in the same way as E3. These results suggest that inactivation of ribosomes is not due to colicin molecules prepared by the standard procedure, but to a modified form of them.
Keywords: ribosomes
Full text
PDF




Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- 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]
- Boon T. Inactivation of ribosomes in vitro by colicin E 3 and its mechanism of action. Proc Natl Acad Sci U S A. 1972 Mar;69(3):549–552. doi: 10.1073/pnas.69.3.549. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Bowman C. M. Inactivation of ribosomes by colicin E3 in vitro: Requirement for 50 S ribosomal subunits. FEBS Lett. 1972 Apr 15;22(1):73–75. doi: 10.1016/0014-5793(72)80222-9. [DOI] [PubMed] [Google Scholar]
- 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]
- Dahlberg A. E., Lund E., Kjeldgaard N. O., Bowman C. M., Nomura M. Colicin E3 induced cleavage of 16S ribosomal ribonucleic acid; blocking effects of certain antibiotics. Biochemistry. 1973 Feb 27;12(5):948–950. doi: 10.1021/bi00729a025. [DOI] [PubMed] [Google Scholar]
- 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]
- Jakes K., Zinder N. D., Boon T. Purification and properties of colicin E3 immunity protein. J Biol Chem. 1974 Jan 25;249(2):438–444. [PubMed] [Google Scholar]
- Konisky J., Nomura M. Interaction of colicins with bacterial cells. II. Specific alteration of Escherichia coli ribosomes induced by colicin E3 in vivo. J Mol Biol. 1967 Jun 14;26(2):181–195. doi: 10.1016/0022-2836(67)90290-2. [DOI] [PubMed] [Google Scholar]
- Maeda A., Nomura M. Interaction of colicins with bacterial cells. I. Studies with radioactive colicins. J Bacteriol. 1966 Feb;91(2):685–694. doi: 10.1128/jb.91.2.685-694.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Meyhack B., Meyhack I., Apirion D. Colicin E3: a unique endoribonuclease. Proc Natl Acad Sci U S A. 1973 Jan;70(1):156–160. doi: 10.1073/pnas.70.1.156. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nomura M. Colicins and related bacteriocins. Annu Rev Microbiol. 1967;21:257–284. doi: 10.1146/annurev.mi.21.100167.001353. [DOI] [PubMed] [Google Scholar]
- Osumi Y., Maeda A. Inactivation of ribosomes by a factor induced by colicin E3. J Biochem. 1972 May;71(5):911–914. doi: 10.1093/oxfordjournals.jbchem.a129844. [DOI] [PubMed] [Google Scholar]
- 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]
- Sidikaro J., Nomura M. E3 immunity substance. A protein from e3-colicinogenic cells that accounts for their immunity to colicin E3. J Biol Chem. 1974 Jan 25;249(2):445–453. [PubMed] [Google Scholar]
- Uchida T., Gill D. M., Pappenheimer A. M., Jr Mutation in the structural gene for diphtheria toxin carried by temperate phage . Nat New Biol. 1971 Sep 1;233(35):8–11. doi: 10.1038/newbio233008a0. [DOI] [PubMed] [Google Scholar]
- Zamir A., Miskin R., Elson D. Inactivation and reactivation of ribosomal subunits: amino acyl-transfer RNA binding activity of the 30 s subunit of Escherichia coli. J Mol Biol. 1971 Sep 14;60(2):347–364. doi: 10.1016/0022-2836(71)90299-3. [DOI] [PubMed] [Google Scholar]