Abstract
The role of aminoacyl-tRNA synthetases in the in vivo synthesis of adenylylated bis(5'-nucleosidyl) tetraphosphates (Ap4N) was studied by measuring the concentration of these nucleotides in Escherichia coli cells overproducing lysyl-, methionyl- phenylalanyl-, or valyl-tRNA synthetase. Overproduction of each aminoacyl-tRNA synthetase (20- to 80-fold) was accompanied by a significant increase in intracellular Ap4N concentration (3- to 14-fold). As expected, non-adenylylated bis(5'-nucleosidyl) tetraphosphate concentration was not changed by synthetase overproduction. It was also verified that overproduction of an inactive methionyl-tRNA synthetase mutant did not modify Ap4N concentration. Ap4N accumulation during heat shock occurred in all strains studied. The increase factor (approximately 50-fold after 1 hr at 48 degrees C) was not changed by overproduction of any of the aminoacyl-tRNA synthetases studied, including that of the heat-inducible form of lysyl-tRNA synthetase from the lysU gene. Together, these results establish that aminoacyl-tRNA synthetases are involved in Ap4N biosynthesis during exponential growth as well as during heat shock.
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Selected References
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- Baker J. C., Jacobson M. K. Alteration of adenyl dinucleotide metabolism by environmental stress. Proc Natl Acad Sci U S A. 1986 Apr;83(8):2350–2352. doi: 10.1073/pnas.83.8.2350. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Baker J. C., Jacobson M. K. Determination of diadenosine 5',5''',-P1,P4-tetraphosphate levels in cultured mammalian cells. Anal Biochem. 1984 Sep;141(2):451–460. doi: 10.1016/0003-2697(84)90070-8. [DOI] [PubMed] [Google Scholar]
- Baril E. F., Coughlin S. A., Zamecnik P. C. 5',5'''-P1, P4 diadenosine tetraphosphate (Ap4A): a putative initiator of DNA replication. Cancer Invest. 1985;3(5):465–471. doi: 10.3109/07357908509039808. [DOI] [PubMed] [Google Scholar]
- Blanquet S., Plateau P., Brevet A. The role of zinc in 5',5'-diadenosine tetraphosphate production by aminoacyl-transfer RNA synthetases. Mol Cell Biochem. 1983;52(1):3–11. doi: 10.1007/BF00230583. [DOI] [PubMed] [Google Scholar]
- Bochner B. R., Lee P. C., Wilson S. W., Cutler C. W., Ames B. N. AppppA and related adenylylated nucleotides are synthesized as a consequence of oxidation stress. Cell. 1984 May;37(1):225–232. doi: 10.1016/0092-8674(84)90318-0. [DOI] [PubMed] [Google Scholar]
- Brevet A., Coste H., Fromant M., Plateau P., Blanquet S. Yeast diadenosine 5',5'''-P1,P4-tetraphosphate alpha,beta-phosphorylase behaves as a dinucleoside tetraphosphate synthetase. Biochemistry. 1987 Jul 28;26(15):4763–4768. doi: 10.1021/bi00389a025. [DOI] [PubMed] [Google Scholar]
- Brevet A., Plateau P., Best-Belpomme M., Blanquet S. Variation of Ap4A and other dinucleoside polyphosphates in stressed Drosophila cells. J Biol Chem. 1985 Dec 15;260(29):15566–15570. [PubMed] [Google Scholar]
- Cameselle J. C., Costas M. J., Günther Sillero M. A., Sillero A. Two low Km hydrolytic activities on dinucleoside 5',5"'-P1,P4-tetraphosphates in rat liver. Characterization as the specific dinucleoside tetraphosphatase and a phosphodiesterase I-like enzyme. J Biol Chem. 1984 Mar 10;259(5):2879–2885. [PubMed] [Google Scholar]
- Cassio D., Waller J. P. Modification of methionyl-tRNA synthetase by proteolytic cleavage and properties of the trypsin-modified enzyme. Eur J Biochem. 1971 May 28;20(2):283–300. doi: 10.1111/j.1432-1033.1971.tb01393.x. [DOI] [PubMed] [Google Scholar]
- Coste H., Brevet A., Plateau P., Blanquet S. Non-adenylylated bis(5'-nucleosidyl) tetraphosphates occur in Saccharomyces cerevisiae and in Escherichia coli and accumulate upon temperature shift or exposure to cadmium. J Biol Chem. 1987 Sep 5;262(25):12096–12103. [PubMed] [Google Scholar]
- Dardel F., Fayat G., Blanquet S. Molecular cloning and primary structure of the Escherichia coli methionyl-tRNA synthetase gene. J Bacteriol. 1984 Dec;160(3):1115–1122. doi: 10.1128/jb.160.3.1115-1122.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ducruix A., Hounwanou N., Reinbolt J., Boulanger Y., Blanquet S. Purification and reversible subunit dissociation of overproduced Escherichia coli phenylalanyl-tRNA synthetase. Biochim Biophys Acta. 1983 Nov 17;741(2):244–250. doi: 10.1016/0167-4781(83)90065-9. [DOI] [PubMed] [Google Scholar]
- Fayat G., Blanquet S., Dessen P., Batelier G., Waller J. P. The molecular weight and subunit composition of phenylalanyl-tRNA synthetase from Escherichia coli K-12. Biochimie. 1974;56(1):35–41. doi: 10.1016/s0300-9084(74)80353-6. [DOI] [PubMed] [Google Scholar]
- Fromant M., Fayat G., Laufer P., Blanquet S. Affinity chromatography of aminoacyl-tRNA syntheses on agarose-hexyl-adenosine-5'-phosphate. Biochimie. 1981 Jun;63(6):541–553. doi: 10.1016/s0300-9084(81)80087-9. [DOI] [PubMed] [Google Scholar]
- Goff S. A., Goldberg A. L. Production of abnormal proteins in E. coli stimulates transcription of lon and other heat shock genes. Cell. 1985 Jun;41(2):587–595. doi: 10.1016/s0092-8674(85)80031-3. [DOI] [PubMed] [Google Scholar]
- Guranowski A., Blanquet S. Phosphorolytic cleavage of diadenosine 5',5'''-P1,P4-tetraphosphate. Properties of homogeneous diadenosine 5',5'''-P1,P4-tetraphosphate alpha, beta-phosphorylase from Saccharomyces cerevisiae. J Biol Chem. 1985 Mar 25;260(6):3542–3547. [PubMed] [Google Scholar]
- Guranowski A., Just G., Holler E., Jakubowski H. Synthesis of diadenosine 5',5'''-P1,P4-tetraphosphate (AppppA) from adenosine 5'-phosphosulfate and adenosine 5'-triphosphate catalyzed by yeast AppppA phosphorylase. Biochemistry. 1988 Apr 19;27(8):2959–2964. doi: 10.1021/bi00408a044. [DOI] [PubMed] [Google Scholar]
- HOAGLAND M. B., KELLER E. B., ZAMECNIK P. C. Enzymatic carboxyl activation of amino acids. J Biol Chem. 1956 Jan;218(1):345–358. [PubMed] [Google Scholar]
- Hilderman R. H., Ortwerth B. J. A preferential role for lysyl-tRNA4 in the synthesis of diadenosine 5',5'''-P1,P4-tetraphosphate by an arginyl-tRNA synthetase-lysyl-tRNA synthetase complex from rat liver. Biochemistry. 1987 Mar 24;26(6):1586–1591. doi: 10.1021/bi00380a015. [DOI] [PubMed] [Google Scholar]
- 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]
- Härtlein M., Frank R., Madern D. Nucleotide sequence of Escherichia coli valyl-tRNA synthetase gene valS. Nucleic Acids Res. 1987 Nov 11;15(21):9081–9082. doi: 10.1093/nar/15.21.9081. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Krauspe R., Parthier B., Wasternack C. Synthesis of diadenosine 5',5'''-P1,P4-tetraphosphate by organellar and cytoplasmic phenylalanyl-tRNA synthetases of Euglena gracilis. FEBS Lett. 1988 Aug 1;235(1-2):275–277. doi: 10.1016/0014-5793(88)81278-x. [DOI] [PubMed] [Google Scholar]
- 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]
- Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
- Lawrence F., Blanquet S., Poiret M., Robert-Gero M., Waller J. P. The mechanism of action of methionyl-tRNA synthetase. 3. Ion requirements and kinetic parameters of the ATP-PPi exchange and methionine-transfer reactions catalyzed by the native and trypsin-modified enzymes. Eur J Biochem. 1973 Jul 2;36(1):234–243. doi: 10.1111/j.1432-1033.1973.tb02905.x. [DOI] [PubMed] [Google Scholar]
- Lee P. C., Bochner B. R., Ames B. N. Diadenosine 5',5"'-P1,P4-tetraphosphate and related adenylylated nucleotides in Salmonella typhimurium. J Biol Chem. 1983 Jun 10;258(11):6827–6834. [PubMed] [Google Scholar]
- Mechulam Y., Blanquet S., Fayat G. Dual level control of the Escherichia coli pheST-himA operon expression. tRNA(Phe)-dependent attenuation and transcriptional operator-repressor control by himA and the SOS network. J Mol Biol. 1987 Oct 5;197(3):453–470. doi: 10.1016/0022-2836(87)90558-4. [DOI] [PubMed] [Google Scholar]
- Mechulam Y., Fromant M., Mellot P., Plateau P., Blanchin-Roland S., Fayat G., Blanquet S. Molecular cloning of the Escherichia coli gene for diadenosine 5',5'''-P1,P4-tetraphosphate pyrophosphohydrolase. J Bacteriol. 1985 Oct;164(1):63–69. doi: 10.1128/jb.164.1.63-69.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mellot P., Mechulam Y., Le Corre D., Blanquet S., Fayat G. Identification of an amino acid region supporting specific methionyl-tRNA synthetase: tRNA recognition. J Mol Biol. 1989 Aug 5;208(3):429–443. doi: 10.1016/0022-2836(89)90507-x. [DOI] [PubMed] [Google Scholar]
- Neidhardt F. C., Bloch P. L., Smith D. F. Culture medium for enterobacteria. J Bacteriol. 1974 Sep;119(3):736–747. doi: 10.1128/jb.119.3.736-747.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Neidhardt F. C., VanBogelen R. A., Vaughn V. The genetics and regulation of heat-shock proteins. Annu Rev Genet. 1984;18:295–329. doi: 10.1146/annurev.ge.18.120184.001455. [DOI] [PubMed] [Google Scholar]
- Ogilvie A. Determination of diadenosine tetraphosphate (Ap4A) levels in subpicomole quantities by a phosphodiesterase luciferin--luciferase coupled assay: application as a specific assay for diadenosine tetraphosphatase. Anal Biochem. 1981 Aug;115(2):302–307. doi: 10.1016/0003-2697(81)90009-9. [DOI] [PubMed] [Google Scholar]
- Plateau P., Blanquet S. Zinc-dependent synthesis of various dinucleoside 5',5' ' '-P1,P3-Tri- or 5'',5' ' '-P1,P4-tetraphosphates by Escherichia coli lysyl-tRNA synthetase. Biochemistry. 1982 Oct 12;21(21):5273–5279. doi: 10.1021/bi00264a024. [DOI] [PubMed] [Google Scholar]
- Plateau P., Fromant M., Brevet A., Gesquière A., Blanquet S. Catabolism of bis(5'-nucleosidyl) oligophosphates in Escherichia coli: metal requirements and substrate specificity of homogeneous diadenosine-5',5'''-P1,P4-tetraphosphate pyrophosphohydrolase. Biochemistry. 1985 Feb 12;24(4):914–922. doi: 10.1021/bi00325a016. [DOI] [PubMed] [Google Scholar]
- 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]
- Prescott M., Milne A. D., McLennan A. G. Characterization of the bis(5'-nucleosidyl) tetraphosphate pyrophosphohydrolase from encysted embryos of the brine shrimp Artemia. Biochem J. 1989 May 1;259(3):831–838. doi: 10.1042/bj2590831. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rapaport E., Zamecnik P. C. Presence of diadenosine 5',5''' -P1, P4-tetraphosphate (Ap4A) in mamalian cells in levels varying widely with proliferative activity of the tissue: a possible positive "pleiotypic activator". Proc Natl Acad Sci U S A. 1976 Nov;73(11):3984–3988. doi: 10.1073/pnas.73.11.3984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Robinson A. K., Barnes L. D. Three diadenosine 5',5''-P1,P4-tetraphosphate hydrolytic enzymes from Physarum polycephalum with differential effects by calcium: a specific dinucleoside polyphosphate pyrophosphohydrolase, a nucleotide pyrophosphatase, and a phosphodiesterase. Arch Biochem Biophys. 1986 Aug 1;248(2):502–515. doi: 10.1016/0003-9861(86)90503-5. [DOI] [PubMed] [Google Scholar]
- Skogman S. G., Nilsson J. Molecular cloning and characterization of the gene for Escherichia coli valyl-tRNA synthetase. Gene. 1984 Oct;30(1-3):219–226. doi: 10.1016/0378-1119(84)90123-9. [DOI] [PubMed] [Google Scholar]
- Traut T. W. Synthesis of hybrid bisnucleoside 5',5"'-P1,P4-tetraphosphates by aminoacyl-tRNA synthetases. Mol Cell Biochem. 1987 May;75(1):15–21. doi: 10.1007/BF00231604. [DOI] [PubMed] [Google Scholar]
- VanBogelen R. A., Vaughn V., Neidhardt F. C. Gene for heat-inducible lysyl-tRNA synthetase (lysU) maps near cadA in Escherichia coli. J Bacteriol. 1983 Feb;153(2):1066–1068. doi: 10.1128/jb.153.2.1066-1068.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Zamecnik P. C., Stephenson M. L., Janeway C. M., Randerath K. Enzymatic synthesis of diadenosine tetraphosphate and diadenosine triphosphate with a purified lysyl-sRNA synthetase. Biochem Biophys Res Commun. 1966 Jul 6;24(1):91–97. doi: 10.1016/0006-291x(66)90415-3. [DOI] [PubMed] [Google Scholar]
- Zamecnik P. Diadenosine 5',5"'-P1,P4-tetraphosphate (Ap4A): its role in cellular metabolism. Anal Biochem. 1983 Oct 1;134(1):1–10. doi: 10.1016/0003-2697(83)90255-5. [DOI] [PubMed] [Google Scholar]