Abstract
Colicin E2 purified by conventional methods contains a tightly bound low-molecular-weight protein, as has been found with purified colicin E3 [Jakes,N.&Zinder,N.D.(1974) Proc. Natl. Acad. Sci. USA 71, 3380-3384]. Such E2 preparations do not cause DNA cleavage in vitro. After separation from the low-molecular-weight protein, colicin E2 retained the original in vivo killing activity, and in addition showed a high activity in vitro in cleaving various DNA molecules, such as a ColE1 hybrid plasmid and DNAs from Escherichia coli, lambda phage, chiX174 phage, and simian virus 40. The low-molecular-weight protein ("E2-immunity protein") specifically prevented this in vitro DNA cleavage reaction, i.e., had an "immunity function." The results demonstrate that colicin E2 itself is a DNA endonuclease and explain the in vivo effects caused by E2 in sensitive cells as well as the mechanism of immunity in E2-colicinogenic cells.
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- Almendinger R., Hager L. P. Role for endonuclease I in the transmission process of colicin E 2 . Nat New Biol. 1972 Feb 16;235(59):199–203. doi: 10.1038/newbio235199a0. [DOI] [PubMed] [Google Scholar]
- Beppu T., Kawabata K., Arima K. Specific inhibition of cell division by colicin E 2 without degradation of deoxyribonucleic acid in a new colicin sensitivity mutant of Escherichia coli. J Bacteriol. 1972 May;110(2):485–493. doi: 10.1128/jb.110.2.485-493.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Clarke L., Carbon J. Biochemical construction and selection of hybrid plasmids containing specific segments of the Escherichia coli genome. Proc Natl Acad Sci U S A. 1975 Nov;72(11):4361–4365. doi: 10.1073/pnas.72.11.4361. [DOI] [PMC free article] [PubMed] [Google Scholar]
- FREDERICQ P. Colicins and colicinogenic factors. Symp Soc Exp Biol. 1958;12:104–122. [PubMed] [Google Scholar]
- GESTELAND R. F., BOEDTKER H. SOME PHYSICAL PROPERTIES OF BACTERIOPHAGE R17 AND ITS RIBONUCLEIC ACID. J Mol Biol. 1964 Apr;8:496–507. doi: 10.1016/s0022-2836(64)80007-3. [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. 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]
- 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]
- LEHMAN I. R., ROUSSOS G. G., PRATT E. A. The deoxyribonucleases of Escherichia coli. II. Purification and properties of a ribonucleic acid-inhibitable endonuclease. J Biol Chem. 1962 Mar;237:819–828. [PubMed] [Google Scholar]
- Maeda A. Existence of some substances which react to colicin E2. Biochim Biophys Acta. 1974 Dec 20;374(3):426–430. doi: 10.1016/0005-2787(74)90264-0. [DOI] [PubMed] [Google Scholar]
- Murray K., Murray N. E. Phage lambda receptor chromosomes for DNA fragments made with restriction endonuclease III of Haemophilus influenzae and restriction endonuclease I of Escherichia coli. J Mol Biol. 1975 Nov 5;98(3):551–564. doi: 10.1016/s0022-2836(75)80086-6. [DOI] [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]
- Nose K., Mizuno D., Ozeki H. Degradation of ribosomal RNA from Escherichia coli induced by colicine E2. Biochim Biophys Acta. 1966 Jun 22;119(3):636–638. doi: 10.1016/0005-2787(66)90142-0. [DOI] [PubMed] [Google Scholar]
- Obinata M., Mizuno D. Mechanism of colicin E2-induced DNA degradation in Escherichia coli. Biochim Biophys Acta. 1970 Feb 18;199(2):330–339. doi: 10.1016/0005-2787(70)90076-6. [DOI] [PubMed] [Google Scholar]
- Ringrose P. S. Interaction between colicin E2 and DNA in vitro. FEBS Lett. 1972 Jun 15;23(2):241–243. doi: 10.1016/0014-5793(72)80351-x. [DOI] [PubMed] [Google Scholar]
- Ringrose P. Sedimentation analysis of DNA degradation products resulting from the action of colicin E2 on Escherichia coli. Biochim Biophys Acta. 1970 Aug 8;213(2):320–334. doi: 10.1016/0005-2787(70)90040-7. [DOI] [PubMed] [Google Scholar]
- Saxe L. S. The action of colicin E2 on supercoiled lambda DNA. I. Experiments in vivo. Biochemistry. 1975 May 20;14(10):2051–2057. doi: 10.1021/bi00681a003. [DOI] [PubMed] [Google Scholar]
- Saxe L. S. The action of colicin E2 on supercoiled lambda DNA.II. Experiments in vitro. Biochemistry. 1975 May 20;14(10):2058–2063. doi: 10.1021/bi00681a004. [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]
- Seto A., Shinozawa T., Maeda A. Inactivation of infectious deoxyribonucleic acid of bacteriophage phi chi X 174 by colicin E2. Biochim Biophys Acta. 1973 Oct 26;324(3):305–308. doi: 10.1016/0005-2787(73)90276-1. [DOI] [PubMed] [Google Scholar]
- Shinnick T. M., Lund E., Smithies O., Blattner F. R. Hybridization of labeled RNA to DNA in agarose gels. Nucleic Acids Res. 1975 Oct;2(10):1911–1929. doi: 10.1093/nar/2.10.1911. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sidikaro J., Masayasu N. In vitro synthesis of the E3 immunity protein directed by Col E3 plasmid deoxyribonucleic acid. J Biol Chem. 1975 Feb 10;250(3):1123–1131. [PubMed] [Google Scholar]
- Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]