Skip to main content
PMC home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1980 May 10;8(9):2023–2037. doi: 10.1093/nar/8.9.2023

Two kinetically distinct tRNAile isoacceptors in Escherichia coli C6.

C L Harris, F Marashi
PMCID: PMC324055  PMID: 6159598

Abstract

The isoleucine acceptance of tRNA from Escherichia coli C6 was previously shown to be influenced by the synthetase level (Marashi, F. and Harris, C.L. 1977. Biochim. Biophys. Acta 477, 84-88). We show here that the increased acceptance observed at higher enzyme levels is accompanied by an increase in the aminoacylation of one tRNAile species. Hence, tRNAile, a minor species at low enzyme levels, is a major isoacceptor after full aminoacylation. The two major isoleucine species have been purified using a combination of BD-cellulose, DEAE-Sephadex A-50 and methylated albumin kieselguhr chromatography. tRNAile (1511 pmoles ile/A260 of tRNA) was found to be slowly acylated, with 2a Vmax one-seventh that observed with tRNAil3le (1475 pmoles ile/A260). Two-dimensional TLC analysis of RNase T2 digests revealed differences in nucleotide content between the purified tRNAs. These results are discussed in terms of the presence of slow and fast tRNAile species in E. coli.

Full text

PDF
2023

Selected References

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

  1. Anderson E. H. Growth Requirements of Virus-Resistant Mutants of Escherichia Coli Strain "B". Proc Natl Acad Sci U S A. 1946 May;32(5):120–128. doi: 10.1073/pnas.32.5.120. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bachmann B. J. Pedigrees of some mutant strains of Escherichia coli K-12. Bacteriol Rev. 1972 Dec;36(4):525–557. doi: 10.1128/br.36.4.525-557.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Baldwin A. N., Berg P. Purification and properties of isoleucyl ribonucleic acid synthetase from Escherichia coli. J Biol Chem. 1966 Feb 25;241(4):831–838. [PubMed] [Google Scholar]
  4. Bonnet J., Ebel J. P. Interpretation of incomplete reactions in tRNA aminoacylation. Aminoacylation of yeast tRNA Val II with yeast valyl-tRNA synthetase. Eur J Biochem. 1972 Dec 4;31(2):335–344. doi: 10.1111/j.1432-1033.1972.tb02538.x. [DOI] [PubMed] [Google Scholar]
  5. Chang S. H., Kuo S., Hawkins E., Miller N. R. The corrected nucleotide sequence of yeast leucine transfer ribonucleic acid. Biochem Biophys Res Commun. 1973 Apr 16;51(4):951–955. doi: 10.1016/0006-291x(73)90019-3. [DOI] [PubMed] [Google Scholar]
  6. Eldred E. W., Schimmel P. R. Investigation of the transfer of amino acid from a transfer ribonucleic acid synthetase-aminoacyl adenylate complex to transfer ribonucleic acid. Biochemistry. 1972 Jan 4;11(1):17–23. doi: 10.1021/bi00751a004. [DOI] [PubMed] [Google Scholar]
  7. Fersht A. R., Kaethner M. M. Mechanism of aminoacylation of tRNA. Proof of the aminoacyl adenylate pathway for the isoleucyl- and tyrosyl-tRNA synthetases from Escherichia coli K12. Biochemistry. 1976 Feb 24;15(4):818–823. doi: 10.1021/bi00649a014. [DOI] [PubMed] [Google Scholar]
  8. Friedman S., Li H. J., Nakanishi K., Van Lear G. 3-(3-amino-3-carboxy-n-propyl)uridine. The structure of the nucleoside in Escherichia coli transfer ribonucleic acid that reacts with phenoxyacetoxysuccinimide. Biochemistry. 1974 Jul 2;13(14):2932–2937. doi: 10.1021/bi00711a024. [DOI] [PubMed] [Google Scholar]
  9. Gillam I., Millward S., Blew D., von Tigerstrom M., Wimmer E., Tener G. M. The separation of soluble ribonucleic acids on benzoylated diethylaminoethylcellulose. Biochemistry. 1967 Oct;6(10):3043–3056. doi: 10.1021/bi00862a011. [DOI] [PubMed] [Google Scholar]
  10. Gusseck D. J. Anomalies in the tRNA-aminoacylation reaction which could lead to misinterpretation of evidence for tRNA changes during development and aging. Mech Ageing Dev. 1974 Nov-Dec;3(5-6):301–309. doi: 10.1016/0047-6374(74)90026-8. [DOI] [PubMed] [Google Scholar]
  11. Harada F., Kimura F., Nishimura S. Primary sequence of tRNA val from Escherichia coli B. I. Oligonucleotide sequences of digests of Escherichia coli tRNA val with RNase T and pancreatic RNase. Biochemistry. 1971 Aug 17;10(17):3269–3277. doi: 10.1021/bi00793a017. [DOI] [PubMed] [Google Scholar]
  12. Harada F., Kimura F., Nishimura S. Primary sequence of tRNA val from Escherichia coli B. I. Oligonucleotide sequences of digests of Escherichia coli tRNA val with RNase T and pancreatic RNase. Biochemistry. 1971 Aug 17;10(17):3269–3277. doi: 10.1021/bi00793a017. [DOI] [PubMed] [Google Scholar]
  13. Harada F., Nishimura S. Purification and characterization of AUA specific isoleucine transfer ribonucleic acid from Escherichia coli B. Biochemistry. 1974 Jan 15;13(2):300–307. doi: 10.1021/bi00699a011. [DOI] [PubMed] [Google Scholar]
  14. Harada F., Nishimura S. Purification and characterization of AUA specific isoleucine transfer ribonucleic acid from Escherichia coli B. Biochemistry. 1974 Jan 15;13(2):300–307. doi: 10.1021/bi00699a011. [DOI] [PubMed] [Google Scholar]
  15. Harris C. L., Marashi F., Titchener E. B. Increased isoleucine acceptance by sulfur-deficient transfer RNA from Escherichia coli. Nucleic Acids Res. 1976 Aug;3(8):2129–2142. doi: 10.1093/nar/3.8.2129. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Harris C. L., Titchener E. B., Cline A. L. Sulfur-deficient transfer ribonucleic acid in a cysteine-requiring, "relaxed" mutant of Escherichia coli. J Bacteriol. 1969 Dec;100(3):1322–1327. doi: 10.1128/jb.100.3.1322-1327.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Ikemura T., Ozeki H. Gross map location of Escherichia coli transfer RNA genes. J Mol Biol. 1977 Dec 5;117(2):419–446. doi: 10.1016/0022-2836(77)90136-x. [DOI] [PubMed] [Google Scholar]
  18. Keith G., Rogg H., Dirheimer G., Menichi B., Heyham T. Post-transcriptional modification of tyrosine tRNA as a function of growth in Bacillus subtilis. FEBS Lett. 1976 Jan 15;61(2):120–123. doi: 10.1016/0014-5793(76)81017-4. [DOI] [PubMed] [Google Scholar]
  19. Kelmers A. D., Novelli G. D., Stulberg M. P. Separation of transfer ribonucleic acids by reverse phase chromatography. J Biol Chem. 1965 Oct;240(10):3979–3983. [PubMed] [Google Scholar]
  20. Kimura-Harada F., Harada F., Nishimura S. The presence of N-[9-(c-D-ribofuranosyl)purin-6-ylcarbamoyl] threonine in isoleucine, threonine and asparagine tRNAs from Escherichia coli. FEBS Lett. 1972 Mar;21(1):71–74. doi: 10.1016/0014-5793(72)80166-2. [DOI] [PubMed] [Google Scholar]
  21. 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]
  22. Lipsett M. N. The isolation of 4-thiouridylic acid from the soluble ribonucleic acid of Escherichia coli. J Biol Chem. 1965 Oct;240(10):3975–3978. [PubMed] [Google Scholar]
  23. Lund E., Dahlberg J. E., Lindahl L., Jaskunas S. R., Dennis P. P., Nomura M. Transfer RNA genes between 16S and 23S rRNA genes in rRNA transcription units of E. coli. Cell. 1976 Feb;7(2):165–177. doi: 10.1016/0092-8674(76)90016-7. [DOI] [PubMed] [Google Scholar]
  24. Lövgren T. N., Pastuszyn A., Loftfield R. B. The mechanism of the aminoacylation of transfer ribonucleic acid: enzyme-product dissociation is not rate limiting. Biochemistry. 1976 Jun 15;15(12):2533–2540. doi: 10.1021/bi00657a007. [DOI] [PubMed] [Google Scholar]
  25. MANDELL J. D., HERSHEY A. D. A fractionating column for analysis of nucleic acids. Anal Biochem. 1960 Jun;1:66–77. doi: 10.1016/0003-2697(60)90020-8. [DOI] [PubMed] [Google Scholar]
  26. Marashi F., Harris C. L. Isoleucyl-tRNA synthetase inactivation and the extent of aminoacylation of tRNAIle from Escherichia coli. Biochim Biophys Acta. 1977 Jul 5;477(1):84–88. doi: 10.1016/0005-2787(77)90162-9. [DOI] [PubMed] [Google Scholar]
  27. Miller J. P., Hussain Z., Schweizer M. P. The involvement of the anticodon adjacent modified nucleoside N-(9-(BETA-D-ribofuranosyl) purine-6-ylcarbamoyl)-threonine in the biological function of E. coli tRNAile. Nucleic Acids Res. 1976 May;3(5):1185–1201. doi: 10.1093/nar/3.5.1185. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Morgan S. D., Söll D. Regulation of the biosynthesis of aminoacid: tRNA ligases and of tRNA. Prog Nucleic Acid Res Mol Biol. 1978;21:181–207. doi: 10.1016/s0079-6603(08)60270-6. [DOI] [PubMed] [Google Scholar]
  29. Nishimura S., Harada F., Narushima U., Seno T. Purification of methionine-, valine-, phenylalanine- and tyrosine-specific tRNA from Escherichia coli. Biochim Biophys Acta. 1967 Jun 20;142(1):133–148. doi: 10.1016/0005-2787(67)90522-9. [DOI] [PubMed] [Google Scholar]
  30. Rao Y. S., Cherayil J. D. Studies on chemical modification of thionucleosides in the transfer ribonucleic acid of Escherichia coli. Biochem J. 1974 Nov;143(2):285–294. doi: 10.1042/bj1430285. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Renaud M., Bollack C., Ebel J. P. Interpretation of incomplete aminoacylation of yeast tRNA-Phe: evidence for a phenylalanyl-tRNA synthetase catalyzed transconformation of tRNA-Phe. Biochimie. 1974;56(9):1203–1209. doi: 10.1016/s0300-9084(74)80012-x. [DOI] [PubMed] [Google Scholar]
  32. Saneyoshi M., Nishimura S. Selective modification of 4-thiouridylate residue in Escherichia coli transfer RNA with cyanogen bromide. Biochim Biophys Acta. 1970 Apr 15;204(2):389–399. doi: 10.1016/0005-2787(70)90158-9. [DOI] [PubMed] [Google Scholar]
  33. Sekiya T., Nishimura S. Sequence of the gene for isoleucine tRNA1 and the surrounding region in a ribosomal RNA operon of Escherichia coli. Nucleic Acids Res. 1979 Feb;6(2):575–592. doi: 10.1093/nar/6.2.575. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Thomale J., Nass G. Alteration of the intracellular concentration of aminoacyl-tRNA synthetases and isoaccepting tRNAs during amino-acid limited growth in Escherichia coli. Eur J Biochem. 1978 Apr 17;85(2):407–418. doi: 10.1111/j.1432-1033.1978.tb12253.x. [DOI] [PubMed] [Google Scholar]
  35. Waters L. C., Shugart L., Yang W. K., Best A. N. Some physical and biological properties of 4-thiouridine- and dihydrouridine-deficient tRNA from chloramphenicol-treated Escherichia coli. Arch Biochem Biophys. 1973 Jun;156(2):780–793. doi: 10.1016/0003-9861(73)90332-9. [DOI] [PubMed] [Google Scholar]
  36. Yarus M., Barrell B. G. The sequence of nucleotides in tRNA Ile from E. coli B. Biochem Biophys Res Commun. 1971 May 21;43(4):729–734. doi: 10.1016/0006-291x(71)90676-0. [DOI] [PubMed] [Google Scholar]
  37. Yegian C. D., Stent G. S. Differential aminoacylation of three species of isoleucine transfer RNA from Escherichia coli. J Mol Biol. 1969 Jan 14;39(1):59–71. doi: 10.1016/0022-2836(69)90333-7. [DOI] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES