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. 1990 Oct;172(10):5686–5689. doi: 10.1128/jb.172.10.5686-5689.1990

Pyrophosphatase is essential for growth of Escherichia coli.

J Chen 1, A Brevet 1, M Fromant 1, F Lévêque 1, J M Schmitter 1, S Blanquet 1, P Plateau 1
PMCID: PMC526883  PMID: 2170325

Abstract

The ppa gene for inorganic pyrophosphatase is essential for the growth of Escherichia coli. A recombinant with a chromosomal ppa::Kanr lesion and a temperature-sensitive replicon with a ppa+ gene showed a temperature-sensitive growth phenotype, and a mutant with the sole ppa+ gene under control of the lac promoter showed inducer-dependent growth. When the lacp-ppa mutant was subcultured without inducer, the pyrophosphatase level decreased, the PPi level increased, and growth stopped. Cellular PPi reached 16 mM about 6 h after growth arrest without loss of cell viability.

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Selected References

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  1. Bourguignon-Van Horen F., Brotcorn A., Caillet-Fauquet P., Diver W. P., Dohet C., Doubleday O. P., Lecomte P., Maenhaut-Michel G., Radman M. Conservation and diversification of genes by mismatch correction and SOS induction. Biochimie. 1982 Aug-Sep;64(8-9):559–564. doi: 10.1016/s0300-9084(82)80087-4. [DOI] [PubMed] [Google Scholar]
  2. Cooperman B. S. The mechanism of action of yeast inorganic pyrophosphatase. Methods Enzymol. 1982;87:526–548. doi: 10.1016/s0076-6879(82)87030-4. [DOI] [PubMed] [Google Scholar]
  3. Coulondre C., Miller J. H. Genetic studies of the lac repressor. III. Additional correlation of mutational sites with specific amino acid residues. J Mol Biol. 1977 Dec 15;117(3):525–567. doi: 10.1016/0022-2836(77)90056-0. [DOI] [PubMed] [Google Scholar]
  4. Csonka L. N., Clark A. J. Construction of an Hfr strain useful for transferring recA mutations between Escherichia coli strains. J Bacteriol. 1980 Jul;143(1):529–530. doi: 10.1128/jb.143.1.529-530.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Hamilton C. M., Aldea M., Washburn B. K., Babitzke P., Kushner S. R. New method for generating deletions and gene replacements in Escherichia coli. J Bacteriol. 1989 Sep;171(9):4617–4622. doi: 10.1128/jb.171.9.4617-4622.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Heinonen J. New radiochemical method for assay of enzymes catalyzing the cleavage of inorganic pyrophosphate. Anal Biochem. 1970 Sep;37(1):32–43. doi: 10.1016/0003-2697(70)90255-1. [DOI] [PubMed] [Google Scholar]
  7. Herbomel P., Ninio J. Fidélité d'une réaction de polymérisation selon la proximité de l'équilibre. C R Seances Acad Sci D. 1980 Nov 24;291(11):881–884. [PubMed] [Google Scholar]
  8. Hirel P. H., Lévêque F., Mellot P., Dardel F., Panvert M., Mechulam Y., Fayat G. Genetic engineering of methionyl-tRNA synthetase: in vitro regeneration of an active synthetase by proteolytic cleavage of a methionyl-tRNA synthetase--beta-galactosidase chimeric protein. Biochimie. 1988 Jun;70(6):773–782. doi: 10.1016/0300-9084(88)90107-1. [DOI] [PubMed] [Google Scholar]
  9. Klemme J. H. Regulation of intracellular pyrophosphatase-activity and conservation of the phosphoanhydride-energy of inorganic pyrophosphate in microbial metabolism. Z Naturforsch C. 1976 Sep-Oct;31(9-10):544–550. doi: 10.1515/znc-1976-9-1011. [DOI] [PubMed] [Google Scholar]
  10. Kohara Y., Akiyama K., Isono K. The physical map of the whole E. coli chromosome: application of a new strategy for rapid analysis and sorting of a large genomic library. Cell. 1987 Jul 31;50(3):495–508. doi: 10.1016/0092-8674(87)90503-4. [DOI] [PubMed] [Google Scholar]
  11. Kukko-Kalske E., Heinonen J. Inorganic pyrophosphate and inorganic pyrophosphatase in Escherichia coli. Int J Biochem. 1985;17(5):575–580. doi: 10.1016/0020-711x(85)90288-5. [DOI] [PubMed] [Google Scholar]
  12. Kukko-Kalske E., Lintunen M., Inen M. K., Lahti R., Heinonen J. Intracellular PPi concentration is not directly dependent on amount of inorganic pyrophosphatase in Escherichia coli K-12 cells. J Bacteriol. 1989 Aug;171(8):4498–4500. doi: 10.1128/jb.171.8.4498-4500.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kukko E., Heinonen J. The intracellular concentration of pyrophosphate in the batch culture of Escherichia coli. Eur J Biochem. 1982 Oct;127(2):347–349. doi: 10.1111/j.1432-1033.1982.tb06878.x. [DOI] [PubMed] [Google Scholar]
  14. Kukko E., Saarento H. Diphosphate concentration does not correlate with the level of inorganic diphosphatase in Escherichia coli. Folia Microbiol (Praha) 1984;29(4):282–287. doi: 10.1007/BF02875958. [DOI] [PubMed] [Google Scholar]
  15. Lahti R., Pitkäranta T., Valve E., Ilta I., Kukko-Kalske E., Heinonen J. Cloning and characterization of the gene encoding inorganic pyrophosphatase of Escherichia coli K-12. J Bacteriol. 1988 Dec;170(12):5901–5907. doi: 10.1128/jb.170.12.5901-5907.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Lecomte P., Doubleday O. P., Radman M. Evidence for an intermediate in DNA synthesis involving pyrophosphate exchange. A possible role in fidelity. J Mol Biol. 1986 Jun 20;189(4):643–652. doi: 10.1016/0022-2836(86)90494-8. [DOI] [PubMed] [Google Scholar]
  17. Nyrén P., Lundin A. Enzymatic method for continuous monitoring of inorganic pyrophosphate synthesis. Anal Biochem. 1985 Dec;151(2):504–509. doi: 10.1016/0003-2697(85)90211-8. [DOI] [PubMed] [Google Scholar]
  18. Plateau P., Fromant M., Kepes F., Blanquet S. Intracellular 5',5'-dinucleoside polyphosphate levels remain constant during the Escherichia coli cell cycle. J Bacteriol. 1987 Jan;169(1):419–422. doi: 10.1128/jb.169.1.419-422.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Sedivy J. M., Daldal F., Fraenkel D. G. Fructose bisphosphatase of Escherichia coli: cloning of the structural gene (fbp) and preparation of a chromosomal deletion. J Bacteriol. 1984 Jun;158(3):1048–1053. doi: 10.1128/jb.158.3.1048-1053.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Simons R. W., Houman F., Kleckner N. Improved single and multicopy lac-based cloning vectors for protein and operon fusions. Gene. 1987;53(1):85–96. doi: 10.1016/0378-1119(87)90095-3. [DOI] [PubMed] [Google Scholar]
  21. Vieira J., Messing J. The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene. 1982 Oct;19(3):259–268. doi: 10.1016/0378-1119(82)90015-4. [DOI] [PubMed] [Google Scholar]
  22. Wallace R. B., Miyada C. G. Oligonucleotide probes for the screening of recombinant DNA libraries. Methods Enzymol. 1987;152:432–442. doi: 10.1016/0076-6879(87)52050-x. [DOI] [PubMed] [Google Scholar]

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