Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1982 Oct;44(1):235–240. doi: 10.1128/jvi.44.1.235-240.1982

Bacteriophage T7 defective in the gene 6 exonuclease promotes site-specific cleavages of T7 DNA in vivo and in vitro.

D D Lee, P D Sadowski
PMCID: PMC256257  PMID: 7143567

Abstract

Site-specific cleavages of intracellular DNA were demonstrated in bacteriophage T7 6am-infected cells. The sites of the cleavages were located at 46.8 and 68.7% (1% of the T7 DNA length = 400 base pairs) from the left end of the T7 genome. These cleavages required the products of genes 3 (endonuclease), 4 (DNA primase), and 5 (DNA polymerase). However, the product of gene 6 (exonuclease) must be absent. Site-specific cleavage was also shown to occur in vitro in extracts of T7 6am-infected cells, although at a different site: 82.8% from the left end of the T7 genome.

Full text

PDF
235

Images in this article

236Image
on p.236
237Image
on p.237
238Image
on p.238
238Image
on p.238
239Image
on p.239

Selected References

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

  1. Bingham A. H., Sharman A. F., Atkinson T. The purification of restriction endonuclease EcoRI by precipitation involving polyethyleneimine. FEBS Lett. 1977 Apr 15;76(2):250–256. doi: 10.1016/0014-5793(77)80162-2. [DOI] [PubMed] [Google Scholar]
  2. Center M. S., Richardson C. C. An endonuclease induced after infection of Escherichia coli with bacteriophage T7. I. Purification and properties of the enzyme. J Biol Chem. 1970 Dec 10;245(23):6285–6291. [PubMed] [Google Scholar]
  3. Center M. S., Studier F. W., Richardson C. C. The structural gene for a T7 endonuclease essential for phage DNA synthesis. Proc Natl Acad Sci U S A. 1970 Jan;65(1):242–248. doi: 10.1073/pnas.65.1.242. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Gellert M., Mizuuchi K., O'Dea M. H., Itoh T., Tomizawa J. I. Nalidixic acid resistance: a second genetic character involved in DNA gyrase activity. Proc Natl Acad Sci U S A. 1977 Nov;74(11):4772–4776. doi: 10.1073/pnas.74.11.4772. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Hobom G., Grosschedl R., Lusky M., Scherer G., Schwarz E., Kössel H. Functional analysis of the replicator structure of lambdoid bacteriophage DNAs. Cold Spring Harb Symp Quant Biol. 1979;43(Pt 1):165–178. doi: 10.1101/sqb.1979.043.01.023. [DOI] [PubMed] [Google Scholar]
  6. Kerr C., Sadowski P. D. Gene 6 exonuclease of bacteriophage T7. I. Purification and properties of the enzyme. J Biol Chem. 1972 Jan 10;247(1):305–310. [PubMed] [Google Scholar]
  7. Kerr C., Sadowski P. D. Gene 6 exonuclease of bacteriophage T7. II. Mechanism of the reaction. J Biol Chem. 1972 Jan 10;247(1):311–318. [PubMed] [Google Scholar]
  8. Kerr C., Sadowski P. D. The involvement of genes 3,4,5 and 6 in genetic recombination in bacteriophage T7. Virology. 1975 May;65(1):281–285. doi: 10.1016/0042-6822(75)90031-8. [DOI] [PubMed] [Google Scholar]
  9. Kikuchi Y., Nash H. A. Nicking-closing activity associated with bacteriophage lambda int gene product. Proc Natl Acad Sci U S A. 1979 Aug;76(8):3760–3764. doi: 10.1073/pnas.76.8.3760. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Lee D., Sadowski P. D. Genetic recombination of bacteriophage T7 in vivo studied by use of a simple physical assay. J Virol. 1981 Dec;40(3):839–847. doi: 10.1128/jvi.40.3.839-847.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Liu L. F., Wang J. C. Interaction between DNA and Escherichia coli DNA topoisomerase I. Formation of complexes between the protein and superhelical and nonsuperhelical duplex DNAs. J Biol Chem. 1979 Nov 10;254(21):11082–11088. [PubMed] [Google Scholar]
  12. Meselson M. S., Radding C. M. A general model for genetic recombination. Proc Natl Acad Sci U S A. 1975 Jan;72(1):358–361. doi: 10.1073/pnas.72.1.358. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Miller R. C., Jr, Lee M. The role of bacteriophage T7 exonuclease (gene 6) in genetic recombination and production of concatemers. J Mol Biol. 1976 Feb 25;101(2):223–234. doi: 10.1016/0022-2836(76)90374-0. [DOI] [PubMed] [Google Scholar]
  14. Panayotatos N., Wells R. D. Cruciform structures in supercoiled DNA. Nature. 1981 Feb 5;289(5797):466–470. doi: 10.1038/289466a0. [DOI] [PubMed] [Google Scholar]
  15. Powling A., Knippers R. Recombination of bacteriophage T7 in vivo. Mol Gen Genet. 1976 Nov 24;149(1):63–71. doi: 10.1007/BF00275961. [DOI] [PubMed] [Google Scholar]
  16. Powling A., Knippers R. Some functions involved in bacteriophage T7 genetic recombination. Mol Gen Genet. 1974;134(2):173–180. doi: 10.1007/BF00268418. [DOI] [PubMed] [Google Scholar]
  17. Roeder G. S., Sadowski P. D. Pathways of recombination of bacteriophage T7 DNA in vitro. Cold Spring Harb Symp Quant Biol. 1979;43(Pt 2):1023–1032. doi: 10.1101/sqb.1979.043.01.111. [DOI] [PubMed] [Google Scholar]
  18. Rosenberg A. H., Simon M. N., Studier F. W., Roberts R. J. Survey and mapping of restriction endonuclease cleavage sites in bacteriophage T7 DNA. J Mol Biol. 1979 Dec 25;135(4):907–915. doi: 10.1016/0022-2836(79)90519-9. [DOI] [PubMed] [Google Scholar]
  19. Sadowski P. D. Bacteriophage T7 endonuclease. I. Properties of the enzyme purified from T7 phage-infected Escherichia coli B. J Biol Chem. 1971 Jan 10;246(1):209–216. [PubMed] [Google Scholar]
  20. Sadowski P. D., Kerr C. Degradation of Escherichia coli B deoxyribonucleic acid after infection with deoxyribonucleic acid-defective amber mutants of bacteriophage T7. J Virol. 1970 Aug;6(2):149–155. doi: 10.1128/jvi.6.2.149-155.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Sadowski P. D., Vetter D. Genetic recombination of bacteriophage T7 DNA in vitro. Proc Natl Acad Sci U S A. 1976 Mar;73(3):692–696. doi: 10.1073/pnas.73.3.692. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Saito H., Tabor S., Tamanoi F., Richardson C. C. Nucleotide sequence of the primary origin of bacteriophage T7 DNA replication: relationship to adjacent genes and regulatory elements. Proc Natl Acad Sci U S A. 1980 Jul;77(7):3917–3921. doi: 10.1073/pnas.77.7.3917. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Shinozaki K., Okazaki T. RNA-linked nascent DNA pieces in T7 phage-infected Escherichia coli cells. I. Role of gene 6 exonuclease in removal of the linked RNA. Mol Gen Genet. 1977 Sep 9;154(3):263–267. doi: 10.1007/BF00571281. [DOI] [PubMed] [Google Scholar]
  24. Shinozaki K., Okazaki T. T7 gene 6 exonuclease has an RNase H activity. Nucleic Acids Res. 1978 Nov;5(11):4245–4261. doi: 10.1093/nar/5.11.4245. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Studier F. W., Rosenberg A. H. Genetic and physical mapping of the late region of bacteriophage T7 DNA by use of cloned fragments of T7 DNA. J Mol Biol. 1981 Dec 15;153(3):503–525. doi: 10.1016/0022-2836(81)90405-8. [DOI] [PubMed] [Google Scholar]
  26. Studier F. W. The genetics and physiology of bacteriophage T7. Virology. 1969 Nov;39(3):562–574. doi: 10.1016/0042-6822(69)90104-4. [DOI] [PubMed] [Google Scholar]
  27. Sugino A., Peebles C. L., Kreuzer K. N., Cozzarelli N. R. Mechanism of action of nalidixic acid: purification of Escherichia coli nalA gene product and its relationship to DNA gyrase and a novel nicking-closing enzyme. Proc Natl Acad Sci U S A. 1977 Nov;74(11):4767–4771. doi: 10.1073/pnas.74.11.4767. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Tsujimoto Y., Ogawa H. EcoRI-sensitive mutation of T7 phage. Mol Gen Genet. 1977 Jan 18;150(2):221–223. doi: 10.1007/BF00695402. [DOI] [PubMed] [Google Scholar]

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

RESOURCES