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. 1972 Jan;109(1):307–314. doi: 10.1128/jb.109.1.307-314.1972

Unusual Valyl-Transfer Ribonucleic Acid Synthetase Mutant of Escherichia coli1

James J Anderson a,2, Frederick C Neidhardt a,3
PMCID: PMC247281  PMID: 4550669

Abstract

Escherichia coli strain NP2907 was isolated as a spontaneous mutant of strain NP29, which possesses a thermolabile valyl-transfer ribonucleic acid (tRNA) synthetase. The valyl-tRNA synthetase of the new mutant, unlike that of its immediate parent, retains enzymatic activity in vitro but differs from the wild-type enzyme in stability and apparent Km for adenosine triphosphate. The new mutant locus, valS-102, cotransduces with pyrB at the same frequency as does the parental locus, valS-1. Cultures of strain NP29 cease growth immediately in any medium when shifted from 30 to 40 C. The new mutant grows normally at 30 C, and upon a shift to 40 C growth quickly accelerates exactly as for normal cells. Exponential growth, however, cannot be sustained at 40 C. At a point characteristic for each medium, growth becomes linear with time. This transition occurs almost immediately in rich media and after 1.5 generations in glucose minimal medium. Net synthesis of valyl-tRNA synthetase ceases in the new mutant as soon as the temperature is raised to 40 C, irrespective of the growth medium. We conclude that it is the amount of valyl-tRNA synthetase activity that limits the rate of growth in the linear phase at 40 C. This property of the mutant makes it possible to evaluate the in vivo efficiency of this enzyme at different growth rates and thereby to determine the concentration that is necessary for a given rate of protein synthesis. The results of our measurements indicate that cells of E. coli growing in minimal medium normally possess a functional excess of valyl-tRNA synthetase with respect to protein synthesis and to repression of threonine deaminase.

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Selected References

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

  1. Anderson J. J., Neidhardt F. C. Growth-linked instability of a mutant valyl-transfer ribonucleic acid synthetase in Escherichia coli. J Bacteriol. 1972 Jan;109(1):315–325. doi: 10.1128/jb.109.1.315-325.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Böck A., Faiman L. E., Neidhardt F. C. Biochemical and genetic characterization of a mutant of Escherichia coli with a temperature-sensitive valyl ribonucleic acid synthetase. J Bacteriol. 1966 Oct;92(4):1076–1082. doi: 10.1128/jb.92.4.1076-1082.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Doolittle W. F., Yanofsky C. Mutants of Escherichia coli with an altered tryptophanyl-transfer ribonucleic acid synthetase. J Bacteriol. 1968 Apr;95(4):1283–1294. doi: 10.1128/jb.95.4.1283-1294.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. EADIE G. S. On the evaluation of the constants Vm and Km in enzyme reactions. Science. 1952 Dec 19;116(3025):688–688. doi: 10.1126/science.116.3025.688. [DOI] [PubMed] [Google Scholar]
  5. EIDLIC L., NEIDHARDT F. C. PROTEIN AND NUCLEIC ACID SYNTHESIS IN TWO MUTANTS OF ESCHERICHIA COLI WITH TEMPERATURE-SENSITIVE AMINOACYL RIBONUCLEIC ACID SYNTHETASES. J Bacteriol. 1965 Mar;89:706–711. doi: 10.1128/jb.89.3.706-711.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. EIDLIC L., NEIDHARDT F. C. ROLE OF VALYL-SRNA SYNTHETASE IN ENZYME REPRESSION. Proc Natl Acad Sci U S A. 1965 Mar;53:539–543. doi: 10.1073/pnas.53.3.539. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. FANGMAN W. L., NEIDHARDT F. C. DEMONSTRATION OF AN ALTERED AMINOACYL RIBONUCLEIC ACID SYNTHETASE IN A MUTANT OF ESCHERICHIA COLI. J Biol Chem. 1964 Jun;239:1839–1843. [PubMed] [Google Scholar]
  8. FRAENKEL D. G., NEIDHARDT F. C. Use of chloramphenicol to study control of RNA synthesis in bacteria. Biochim Biophys Acta. 1961 Oct 14;53:96–110. doi: 10.1016/0006-3002(61)90797-1. [DOI] [PubMed] [Google Scholar]
  9. Freundlich M. Valyl-Transfer RNA: Role in Repression of the Isoleucine-Valine Enzymes in Escherichia coli. Science. 1967 Aug 18;157(3790):823–825. doi: 10.1126/science.157.3790.823-a. [DOI] [PubMed] [Google Scholar]
  10. Hirshfield I. N., Horn P. C., Hopwood D. A., Maas W. K., DeDeken R. Studies on the mechanism of repression of arginine biosynthesis in Escherichia coli. 3. Repression of enzymes of arginine biosynthesis in arginyl-tRNA synthetase mutants. J Mol Biol. 1968 Jul 14;35(1):83–93. doi: 10.1016/s0022-2836(68)80038-5. [DOI] [PubMed] [Google Scholar]
  11. Kaplan S., Anderson D. Selection of temperature-sensitive activating enzyme mutants in Escherichia coli. J Bacteriol. 1968 Mar;95(3):991–997. doi: 10.1128/jb.95.3.991-997.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. LENNOX E. S. Transduction of linked genetic characters of the host by bacteriophage P1. Virology. 1955 Jul;1(2):190–206. doi: 10.1016/0042-6822(55)90016-7. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. Nass G. Regulation of histidine biosynthetic enzymes in a mutant of Escherichia coli with an altered histidyl-tRNA synthetase. Mol Gen Genet. 1967;100(2):216–224. doi: 10.1007/BF00333608. [DOI] [PubMed] [Google Scholar]
  15. Neidhardt F. C. Roles of amino acid activating enzymes in cellular physiology. Bacteriol Rev. 1966 Dec;30(4):701–719. doi: 10.1128/br.30.4.701-719.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Tingle M. A., Neidhardt F. C. Mapping of a structural gene for valyl-transfer ribonucleic acid synthetase in Escherichia coli by transduction. J Bacteriol. 1969 May;98(2):837–839. doi: 10.1128/jb.98.2.837-839.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. UMBARGER H. E., BROWN B. Isoleucine and valine metabolism in Escherichia coli. VII. A negative feedback mechanism controlling isoleucine biosynthesis. J Biol Chem. 1958 Aug;233(2):415–420. [PubMed] [Google Scholar]
  18. Williams L. S., Freundlich M. Role of valine transfer RNA in control of RNA synthesis in Escherichia coli. Biochim Biophys Acta. 1969 Apr 22;179(2):515–517. doi: 10.1016/0005-2787(69)90064-1. [DOI] [PubMed] [Google Scholar]
  19. Williams L. S., Neidhardt F. C. Synthesis and inactivation of aminoacyl-transfer RNA synthetases during growth of Escherichia coli. J Mol Biol. 1969 Aug 14;43(3):529–550. doi: 10.1016/0022-2836(69)90357-x. [DOI] [PubMed] [Google Scholar]
  20. Yaniv M., Gros F. Studies on valyl-tRNA synthetase and tRNA from Escherichia coli. 3. Valyl-tRNA synthetases from thermosensitive mutants of Escherichia coli. J Mol Biol. 1969 Aug 28;44(1):31–45. doi: 10.1016/0022-2836(69)90403-3. [DOI] [PubMed] [Google Scholar]

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