Skip to main content
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1979 May;138(2):530–534. doi: 10.1128/jb.138.2.530-534.1979

Escherichia coli mutant containing a large deletion from relA to argA.

A G Atherly
PMCID: PMC218209  PMID: 374393

Abstract

A mutant of Escherichia coli has been isolated that contains a large deletion (about 3 X 10(7) daltons of deoxyribonucleic acid) encompassing argA, fuc, and relA. This mutant strain (AA-787) is also cold sensitive for growth at 18 degrees C. Strain AA-787 was obtained fortuitously as a cold-sensitive pseudorevertant of a strain having a heat-sensitive peptidyl-transfer ribonucleic acid hydrolase. Genetic analysis, using transduction and interrupted mating, showed the cold sensitivity mutation to be located adjacent to relA. Further analysis demonstrated loss of relA, fuc, and argA gene functions but retention of eno and recB, closely linked genes adjacent to relA and argA, respectively. Unusually high cotransduction of flanking markers (cysC and thyA) indicated loss of approximately 1 min of the E. coli genetic map in strain AA-787. Guanosine 3'-diphosphate 5'-diphosphate (ppGpp) was synthetized in mutant strain AA-787 at basal levels, and ppGpp synthesis was stimulated by carbon-source downshift. No ppGpp synthesis could be obtained using ribosomes isolated from strain AA-787. These findings, taken together, show that deletion of relA in E. coli does not completely abolish ppGpp synthesis and suggests that another enzyme system must also be responsible for ppGpp synthesis.

Full text

PDF
530

Selected References

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

  1. Atherly A. G., Menninger J. R. Mutant E. coli strain with temperature sensitive peptidyl-transfer RNA hydrolase. Nat New Biol. 1972 Dec 20;240(103):245–246. doi: 10.1038/newbio240245a0. [DOI] [PubMed] [Google Scholar]
  2. Atherly A. G., Suchanek M. C. Characterization of mutants of Escherichia coli temperature-sensitive for ribonucleic acid regulation: an unusual phenotype associated with a phenylalanyl transfer ribonucleic acid synthetase mutant. J Bacteriol. 1971 Nov;108(2):627–638. doi: 10.1128/jb.108.2.627-638.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. BOREK E., RYAN A., ROCKENBACH J. Nucleic acid metabolism in relation to the lysogenic phenomenon. J Bacteriol. 1955 Apr;69(4):460–467. doi: 10.1128/jb.69.4.460-467.1955. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bachmann B. J., Low K. B., Taylor A. L. Recalibrated linkage map of Escherichia coli K-12. Bacteriol Rev. 1976 Mar;40(1):116–167. doi: 10.1128/br.40.1.116-167.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Block R., Haseltine W. A. Thermolability of the stringent factor in rel mutants of Escherichia coli. J Mol Biol. 1973 Jul 15;77(4):625–629. doi: 10.1016/0022-2836(73)90228-3. [DOI] [PubMed] [Google Scholar]
  6. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  7. Cashel M. Regulation of bacterial ppGpp and pppGpp. Annu Rev Microbiol. 1975;29:301–318. doi: 10.1146/annurev.mi.29.100175.001505. [DOI] [PubMed] [Google Scholar]
  8. Cashel M. The control of ribonucleic acid synthesis in Escherichia coli. IV. Relevance of unusual phosphorylated compounds from amino acid-starved stringent strains. J Biol Chem. 1969 Jun 25;244(12):3133–3141. [PubMed] [Google Scholar]
  9. Cochran J. W., Byrne R. W. Isolation and properties of a ribosome-bound factor required for ppGpp and ppGpp synthesis in Escherichia coli. J Biol Chem. 1974 Jan 25;249(2):353–360. [PubMed] [Google Scholar]
  10. Diderichsen B., Fiil N. P., Lavallé R. Genetics of the relB locus in Escherichia coli. J Bacteriol. 1977 Jul;131(1):30–33. doi: 10.1128/jb.131.1.30-33.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Fiil N., Friesen J. D. Isolation of "relaxed" mutants of Escherichia coli. J Bacteriol. 1968 Feb;95(2):729–731. doi: 10.1128/jb.95.2.729-731.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Fill N. A functional analysis of the rel gene in Escherichia coli. J Mol Biol. 1969 Oct 28;45(2):195–203. doi: 10.1016/0022-2836(69)90099-0. [DOI] [PubMed] [Google Scholar]
  13. Friesen J. D., An G., Fiil N. P. Nonsense and insertion mutants in the relA gene of E. coli: cloning relA. Cell. 1978 Dec;15(4):1187–1197. doi: 10.1016/0092-8674(78)90045-4. [DOI] [PubMed] [Google Scholar]
  14. Friesen J. D., Fiil N. P., Parker J. M., Haseltine W. A. A new relaxed mutant of Escherichia coli with an altered 50S ribosomal subunit. Proc Natl Acad Sci U S A. 1974 Sep;71(9):3465–3469. doi: 10.1073/pnas.71.9.3465. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hansen M. T., Pato M. L., Molin S., Fill N. P., von Meyenburg K. Simple downshift and resulting lack of correlation between ppGpp pool size and ribonucleic acid accumulation. J Bacteriol. 1975 May;122(2):585–591. doi: 10.1128/jb.122.2.585-591.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Haseltine W. A., Block R. Synthesis of guanosine tetra- and pentaphosphate requires the presence of a codon-specific, uncharged transfer ribonucleic acid in the acceptor site of ribosomes. Proc Natl Acad Sci U S A. 1973 May;70(5):1564–1568. doi: 10.1073/pnas.70.5.1564. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Jenkins S. J., Sparkes C. A., Jones-Mortimer M. C. A gene involved in lysine excretion in Escherichia coli K12. Heredity (Edinb) 1974 Jun;32(3):409–412. doi: 10.1038/hdy.1974.51. [DOI] [PubMed] [Google Scholar]
  18. Menninger J. R., Walker C., Tan P. F. Studies on the metabolic role of peptidyl-tRNA hydrolase. I. Properties of a mutant E. coli with temperature-sensitive peptidyl-tRNA hydrolase. Mol Gen Genet. 1973 Mar 19;121(4):307–324. doi: 10.1007/BF00433230. [DOI] [PubMed] [Google Scholar]
  19. Pao C. C., Gallant J. A gene involved in the metabolic control of ppGpp synthesis. Mol Gen Genet. 1978 Jan 17;158(3):271–277. doi: 10.1007/BF00267198. [DOI] [PubMed] [Google Scholar]
  20. Pedersen F. S., Lund E., Kjeldgaard N. O. Codon specific, tRNA dependent in vitro synthesis of ppGpp and pppGpp. Nat New Biol. 1973 May 2;243(122):13–15. [PubMed] [Google Scholar]
  21. Spring T. G., Wold F. The purification and characterization of Escherichia coli enolase. J Biol Chem. 1971 Nov 25;246(22):6797–6802. [PubMed] [Google Scholar]
  22. Sy J. A ribosome-independent, soluble stringent factor-like enzyme isolated from a Bacillus brevis. Biochemistry. 1976 Feb 10;15(3):606–609. doi: 10.1021/bi00648a024. [DOI] [PubMed] [Google Scholar]
  23. Sy J. In vitro degradation of guanosine 5'-diphosphate, 3'-diphosphate. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5529–5533. doi: 10.1073/pnas.74.12.5529. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Wu T. T. A model for three-point analysis of random general transduction. Genetics. 1966 Aug;54(2):405–410. doi: 10.1093/genetics/54.2.405. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Zabos P., Bauer P., Schlotthauer J., Horváth I. Stringent factor-independent synthesis of pppGpp in Escherichia coli strains. FEBS Lett. 1976 Apr 15;64(1):107–110. doi: 10.1016/0014-5793(76)80261-x. [DOI] [PubMed] [Google Scholar]

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

RESOURCES