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. 1989 Aug;8(8):2411–2415. doi: 10.1002/j.1460-2075.1989.tb08371.x

In vitro reconstitution of anticodon nuclease from components encoded by phage T4 and Escherichia coli CTr5X.

M Amitsur 1, I Morad 1, G Kaufmann 1
PMCID: PMC401184  PMID: 2676520

Abstract

During phage T4 infection of Escherichia coli strains containing the prr locus the host tRNALys undergoes cleavage-ligation in reactions catalyzed by anticodon nuclease, polynucleotide kinase and RNA ligase. Known genetic determinants of anticodon nuclease are prr, which restricts T4 mutants lacking polynucleotide kinase or RNA ligase, and stp, the T4 suppressor of prr encoded restriction. The present communication describes an in vitro anticodon nuclease assay in which the specific cleavage of tRNALys is driven by an extract from E. coli prrr (restrictive) cells infected by phage T4. The in vitro anticodon nuclease reaction requires factor(s) encoded by prr, is stimulated by a synthetic Stp polypeptide and appears to require additional T4 induced factor(s) distinct from Stp.

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Selected References

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  1. Amitsur M., Levitz R., Kaufmann G. Bacteriophage T4 anticodon nuclease, polynucleotide kinase and RNA ligase reprocess the host lysine tRNA. EMBO J. 1987 Aug;6(8):2499–2503. doi: 10.1002/j.1460-2075.1987.tb02532.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Chambliss G. H., Henkin T. M., Leventhal J. M. Bacterial in vitro protein-synthesizing systems. Methods Enzymol. 1983;101:598–605. doi: 10.1016/0076-6879(83)01040-x. [DOI] [PubMed] [Google Scholar]
  3. Chapman D., Morad I., Kaufmann G., Gait M. J., Jorissen L., Snyder L. Nucleotide and deduced amino acid sequence of stp: the bacteriophage T4 anticodon nuclease gene. J Mol Biol. 1988 Jan 20;199(2):373–377. doi: 10.1016/0022-2836(88)90320-8. [DOI] [PubMed] [Google Scholar]
  4. David M., Borasio G. D., Kaufmann G. Bacteriophage T4-induced anticodon-loop nuclease detected in a host strain restrictive to RNA ligase mutants. Proc Natl Acad Sci U S A. 1982 Dec;79(23):7097–7101. doi: 10.1073/pnas.79.23.7097. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. David M., Borasio G. D., Kaufmann G. T4 bacteriophage-coded polynucleotide kinase and RNA ligase are involved in host tRNA alteration or repair. Virology. 1982 Dec;123(2):480–483. doi: 10.1016/0042-6822(82)90284-7. [DOI] [PubMed] [Google Scholar]
  6. Depew R. E., Cozzarelli N. R. Genetics and physiology of bacteriophage T4 3'-phosphatase: evidence for involvement of the enzyme in T4 DNA metabolism. J Virol. 1974 Apr;13(4):888–897. doi: 10.1128/jvi.13.4.888-897.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Depew R. E., Snopek T. J., Cozzarelli N. R. Characterization of a new class of deletions of the D region of the bacteriophage T4 genome. Virology. 1975 Mar;64(1):144–145. doi: 10.1016/0042-6822(75)90086-0. [DOI] [PubMed] [Google Scholar]
  8. GAREN A., OTSUJI N. ISOLATION OF A PROTEIN SPECIFIED BY A REGULATOR GENE. J Mol Biol. 1964 Jun;8:841–852. doi: 10.1016/s0022-2836(64)80165-0. [DOI] [PubMed] [Google Scholar]
  9. Jabbar M. A., Snyder L. Genetic and physiological studies of an Escherichia coli locus that restricts polynucleotide kinase- and RNA ligase-deficient mutants of bacteriophage T4. J Virol. 1984 Aug;51(2):522–529. doi: 10.1128/jvi.51.2.522-529.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kaufmann G., Amitsur M. Host transfer RNA cleavage and reunion in T4-infected Escherichia coli CTr5x. Nucleic Acids Res. 1985 Jun 25;13(12):4333–4341. doi: 10.1093/nar/13.12.4333. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kaufmann G., David M., Borasio G. D., Teichmann A., Paz A., Amitsur M. Phage and host genetic determinants of the specific anticodon loop cleavages in bacteriophage T4-infected Escherichia coli CTr5X. J Mol Biol. 1986 Mar 5;188(1):15–22. doi: 10.1016/0022-2836(86)90476-6. [DOI] [PubMed] [Google Scholar]
  12. Runnels J. M., Soltis D., Hey T., Snyder L. Genetic and physiological studies of the role of the RNA ligase of bacteriophage T4. J Mol Biol. 1982 Jan 15;154(2):273–286. doi: 10.1016/0022-2836(82)90064-x. [DOI] [PubMed] [Google Scholar]
  13. Sirotkin K., Cooley W., Runnels J., Snyder L. R. A role in true-late gene expression for the T4 bacteriophage 5' polynucleotide kinase 3' phosphatase. J Mol Biol. 1978 Aug 5;123(2):221–233. doi: 10.1016/0022-2836(78)90322-4. [DOI] [PubMed] [Google Scholar]
  14. Snyder L., Gold L., Kutter E. A gene of bacteriophage T4 whose product prevents true late transcription on cytosine-containing T4 DNA. Proc Natl Acad Sci U S A. 1976 Sep;73(9):3098–3102. doi: 10.1073/pnas.73.9.3098. [DOI] [PMC free article] [PubMed] [Google Scholar]

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