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. 1974 Mar;71(3):1004–1007. doi: 10.1073/pnas.71.3.1004

A Mutant of Escherichia coli Defective in Leucyl, Phenylalanyl-tRNA-Protein Transferase*

R L Soffer 1, M Savage 1
PMCID: PMC388146  PMID: 4595564

Abstract

A mutant of E. coli that lacks leucyl,-phenylalanyl-tRNA-protein transferase (EC 2.3.2.6) has been isolated. Ability to produce the two activities could be introduced into the mutant from an F′ strain whose episome contains genetic material located between 45 and 54 min on the E. coli chromosome. When grown into stationary phase and resuspended in minimal medium with glycerol, the mutant exhibited a marked lag before resuming growth. Revertants that did not show this lag were selected and were found to have regained both transfer activities. Extracts of wild-type, mutant, and revertant strains were compared as acceptors for the enzymatic transfer of radioactive phenylalanine. Analysis of the labeled polypeptides by disc gel electrophoresis indicated that certain potential acceptors may be preferentially acylated in vivo.

These data provide genetic confirmation that the same enzyme protein catalyzes the transfer of leucine and phenylalanine and suggest that leucyl,phenylalanyl-tRNA-protein transferase is involved in a growth regulatory mechanism.

Keywords: regulation of growth, post-translational modification

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Selected References

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

  1. Burgess R. R. Separation and characterization of the subunits of ribonucleic acid polymerase. J Biol Chem. 1969 Nov 25;244(22):6168–6176. [PubMed] [Google Scholar]
  2. Godson G. N., Sinsheimer R. L. Lysis of Escherichia coli with a neutral detergent. Biochim Biophys Acta. 1967 Dec 19;149(2):476–488. doi: 10.1016/0005-2787(67)90175-x. [DOI] [PubMed] [Google Scholar]
  3. HERZENBERG L. A. Studies on the induction of beta-galactosidase in a cryptic strain of Escherichia coli. Biochim Biophys Acta. 1959 Feb;31(2):525–538. doi: 10.1016/0006-3002(59)90029-0. [DOI] [PubMed] [Google Scholar]
  4. Jacobson A., Gillespie D. Metabolic events occurring during recovery from prolonged glucose starvation in Escherichia coli. J Bacteriol. 1968 Mar;95(3):1030–1039. doi: 10.1128/jb.95.3.1030-1039.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. KAJI A., KAJI H., NOVELLI G. D. SOLUBLE AMINO ACID-INCORPORATING SYSTEM. I. PREPARATION OF THE SYSTEM AND NATURE OF THE REACTION. J Biol Chem. 1965 Mar;240:1185–1191. [PubMed] [Google Scholar]
  6. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  7. Leibowitz M. J., Soffer R. L. Enzymatic modification of proteins. 3. Purification and properties of a leucyl, phenylalanyl transfer ribonucleic acid protein transferase from Escherichia coli. J Biol Chem. 1970 Apr 25;245(8):2066–2073. [PubMed] [Google Scholar]
  8. Leibowitz M. J., Soffer R. L. Enzymatic modification of proteins. VII. Substrate specificity of leucyl,phenylalanyl-transfer ribonucleic acid-protein transferase. J Biol Chem. 1971 Sep 10;246(17):5207–5212. [PubMed] [Google Scholar]
  9. Leibowitz M. J., Soffer R. L. Modification of a specific ribosomal protein catalyzed by leucyl, phenylalanyl-tRNA: protein transferase. Proc Natl Acad Sci U S A. 1971 Aug;68(8):1866–1869. doi: 10.1073/pnas.68.8.1866. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. MANS R. J., NOVELLI G. D. A convenient, rapid and sensitive method for measuring the incorporation of radioactive amino acids into protein. Biochem Biophys Res Commun. 1960 Nov;3:540–543. doi: 10.1016/0006-291x(60)90171-6. [DOI] [PubMed] [Google Scholar]
  11. Momose K., Kaji A. Soluble amino acid-incorporating system. 3. Further studies on the product and its relation to the ribosomal system for incorporation. J Biol Chem. 1966 Jul 25;241(14):3294–3307. [PubMed] [Google Scholar]
  12. NIRENBERG M. W., MATTHAEI J. H. The dependence of cell-free protein synthesis in E. coli upon naturally occurring or synthetic polyribonucleotides. Proc Natl Acad Sci U S A. 1961 Oct 15;47:1588–1602. doi: 10.1073/pnas.47.10.1588. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Soffer R. L. Peptide acceptors in the leucine, phenylalanine transfer reaction. J Biol Chem. 1973 Dec 25;248(24):8424–8428. [PubMed] [Google Scholar]
  14. Taylor A. L. Current linkage map of Escherichia coli. Bacteriol Rev. 1970 Jun;34(2):155–175. doi: 10.1128/br.34.2.155-175.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]

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