Abstract
The Old protein of bacteriophage P2 is responsible for interference with the growth of phage lambda and for killing of recBC mutant Escherichia coli. We have purified Old fused to the maltose-binding protein to 95% purity and characterized its enzymatic properties. The Old protein fused to maltose-binding protein has exonuclease activity on double-stranded DNA as well as nuclease activity on single-stranded DNA and RNA. The direction of digestion of double-stranded DNA is from 5' to 3', and digestion initiates at either the 5'-phosphoryl or 5'-hydroxyl terminus. The nuclease is active on nicked circular DNA, degrades DNA in a processive manner, and releases 5'-phosphoryl mononucleotides.
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- Brégégère F. Bacteriophage P2-lambda interference. II. Effects on the host under the control of lambda genes O and P. J Mol Biol. 1976 Jun 25;104(2):411–420. doi: 10.1016/0022-2836(76)90279-5. [DOI] [PubMed] [Google Scholar]
- Enquist L. W., Skalka A. Replication of bacteriophage lambda DNA dependent on the function of host and viral genes. I. Interaction of red, gam and rec. J Mol Biol. 1973 Apr 5;75(2):185–212. doi: 10.1016/0022-2836(73)90016-8. [DOI] [PubMed] [Google Scholar]
- Finkel S., Halling C., Calendar R. Selection of lambda Spi- transducing phages using the P2 old gene cloned onto a plasmid. Gene. 1986;46(1):65–69. doi: 10.1016/0378-1119(86)90167-8. [DOI] [PubMed] [Google Scholar]
- Gillen J. R., Karu A. E., Nagaishi H., Clark A. J. Characterization of the deoxyribonuclease determined by lambda reverse as exonuclease VIII of Escherichia coli. J Mol Biol. 1977 Jun 15;113(1):27–41. doi: 10.1016/0022-2836(77)90039-0. [DOI] [PubMed] [Google Scholar]
- Goldmark P. J., Linn S. Purification and properties of the recBC DNase of Escherichia coli K-12. J Biol Chem. 1972 Mar 25;247(6):1849–1860. [PubMed] [Google Scholar]
- Haggård-Ljungquist E., Barreiro V., Calendar R., Kurnit D. M., Cheng H. The P2 phage old gene: sequence, transcription and translational control. Gene. 1989 Dec 21;85(1):25–33. doi: 10.1016/0378-1119(89)90460-5. [DOI] [PubMed] [Google Scholar]
- Joseph J. W., Kolodner R. Exonuclease VIII of Escherichia coli. I. Purification and physical properties. J Biol Chem. 1983 Sep 10;258(17):10411–10417. [PubMed] [Google Scholar]
- Joseph J. W., Kolodner R. Exonuclease VIII of Escherichia coli. II. Mechanism of action. J Biol Chem. 1983 Sep 10;258(17):10418–10424. [PubMed] [Google Scholar]
- Kahn M. L., Ziermann R., Dehò G., Ow D. W., Sunshine M. G., Calendar R. Bacteriophage P2 and P4. Methods Enzymol. 1991;204:264–280. doi: 10.1016/0076-6879(91)04013-e. [DOI] [PubMed] [Google Scholar]
- Koonin E. V., Gorbalenya A. E. The superfamily of UvrA-related ATPases includes three more subunits of putative ATP-dependent nucleases. Protein Seq Data Anal. 1992;5(1):43–45. [PubMed] [Google Scholar]
- Kunkel T. A., Roberts J. D., Zakour R. A. Rapid and efficient site-specific mutagenesis without phenotypic selection. Methods Enzymol. 1987;154:367–382. doi: 10.1016/0076-6879(87)54085-x. [DOI] [PubMed] [Google Scholar]
- Kushner S. R., Nagaishi H., Clark A. J. Indirect suppression of recB and recC mutations by exonuclease I deficiency. Proc Natl Acad Sci U S A. 1972 Jun;69(6):1366–1370. doi: 10.1073/pnas.69.6.1366. [DOI] [PMC free article] [PubMed] [Google Scholar]
- LEHMAN I. R. The deoxyribonucleases of Escherichia coli. I. Purification and properties of a phosphodiesterase. J Biol Chem. 1960 May;235:1479–1487. [PubMed] [Google Scholar]
- Lindahl G., Sironi G., Bialy H., Calendar R. Bacteriophage lambda; abortive infection of bacteria lysogenic for phage P2. Proc Natl Acad Sci U S A. 1970 Jul;66(3):587–594. doi: 10.1073/pnas.66.3.587. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Linderoth N. A., Ziermann R., Haggård-Ljungquist E., Christie G. E., Calendar R. Nucleotide sequence of the DNA packaging and capsid synthesis genes of bacteriophage P2. Nucleic Acids Res. 1991 Dec;19(25):7207–7214. doi: 10.1093/nar/19.25.7207. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Little J. W. An exonuclease induced by bacteriophage lambda. II. Nature of the enzymatic reaction. J Biol Chem. 1967 Feb 25;242(4):679–686. [PubMed] [Google Scholar]
- Mahajan S. K., Chu C. C., Willis D. K., Templin A., Clark A. J. Physical analysis of spontaneous and mutagen-induced mutants of Escherichia coli K-12 expressing DNA exonuclease VIII activity. Genetics. 1990 Jun;125(2):261–273. doi: 10.1093/genetics/125.2.261. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Masamune Y., Fleischman R. A., Richardson C. C. Enzymatic removal and replacement of nucleotides at single strand breaks in deoxyribonucleic acid. J Biol Chem. 1971 Apr 25;246(8):2680–2691. [PubMed] [Google Scholar]
- Park C. H., Sancar A. Formation of a ternary complex by human XPA, ERCC1, and ERCC4(XPF) excision repair proteins. Proc Natl Acad Sci U S A. 1994 May 24;91(11):5017–5021. doi: 10.1073/pnas.91.11.5017. [DOI] [PMC free article] [PubMed] [Google Scholar]
- SULKOWSKI E., BJORK W., LASKOWSKI M., Sr A specific and nonspecific alkaline monophosphatase in the venom of Bothrops atrox and their occurrence in the purified venom phosphodiesterase. J Biol Chem. 1963 Jul;238:2477–2486. [PubMed] [Google Scholar]
- Sironi G., Bialy H., Lozeron H. A., Calendar R. Bacteriophage P2: interaction with phage lambda and with recombination-deficient bacteria. Virology. 1971 Nov;46(2):387–396. doi: 10.1016/0042-6822(71)90040-7. [DOI] [PubMed] [Google Scholar]
- Sironi G. Mutants of Escherichia coli unable to be lysogenized by the temperate bacteriophage P2. Virology. 1969 Feb;37(2):163–176. doi: 10.1016/0042-6822(69)90196-2. [DOI] [PubMed] [Google Scholar]
- Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]