Skip to main content
NCBI home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1978 Oct;5(10):3665–3677. doi: 10.1093/nar/5.10.3665

Reactions at the termini of tRNA with T4 RNA ligase.

A G Bruce, O C Uhlenbeck
PMCID: PMC342702  PMID: 724498

Abstract

T4 RNA ligase will catalyze the addition of nucleoside 3', 5'-bisphosphates onto the 3' terminus of tRNA resulting in tRNA molecule one nucleotide longer with a 3' terminal phosphate. Under appropriate conditions the reaction is quantitative and, if high specific radioactivity bisphosphates are used, it provides an efficient means for in vitro labeling of tRNA. Although the 3' terminal hydroxyl is a good acceptor, the 5' terminal phosphate in most tRNA's is not an effective donor in the RNA ligase reaction. This poor reactivity is due to the secondary structure of the 5' terminal nucleotide. If E. Coli tRNAf Met is used, the 5' phosphate is reactive and the major product with RNA ligase is the cyclic tRNA.

Full text

PDF
3665

Images in this article

3668Image
on p.3668
3674Image
on p.3674

Selected References

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

  1. Barrio J. R., Barrio M. C., Leonard N. J., England T. E., Uhlenbeck O. C. Synthesis of modified nucleoside 3',5'-bisphosphates and their incorporation into oligoribonucleotides with T4 RNA ligase. Biochemistry. 1978 May 30;17(11):2077–2081. doi: 10.1021/bi00604a009. [DOI] [PubMed] [Google Scholar]
  2. CANNON M. INSTABILITY OF THE TERMINAL ADENOSINE OF SOLUBLE RIBONUCLEIC ACID OF ESCHERICHIA COLI IN VIVO. Biochim Biophys Acta. 1964 May 18;87:154–156. doi: 10.1016/0926-6550(64)90056-8. [DOI] [PubMed] [Google Scholar]
  3. Cranston J. W., Silber R., Malathi V. G., Hurwitz J. Studies on ribonucleic acid ligase. Characterization of an adenosine triphosphate-inorganic pyrophosphate exchange reaction and demonstration of an enzyme-adenylate complex with T4 bacteriophage-induced enzyme. J Biol Chem. 1974 Dec 10;249(23):7447–7456. [PubMed] [Google Scholar]
  4. Dahlberg A. E., Dingman C. W., Peacock A. C. Electrophoretic characterization of bacterial polyribosomes in agarose-acrylamide composite gels. J Mol Biol. 1969 Apr 14;41(1):139–147. doi: 10.1016/0022-2836(69)90131-4. [DOI] [PubMed] [Google Scholar]
  5. Dale R. M., Martin E., Livingston D. C., Ward D. C. Direct covalent mercuration of nucleotides and polynucleotides. Biochemistry. 1975 Jun 3;14(11):2447–2457. doi: 10.1021/bi00682a027. [DOI] [PubMed] [Google Scholar]
  6. Dale R. M., Ward D. C., Livingston D. C., Martin E. Conversion of covalently mercurated nucleic acids to tritiated and halogenated derivatives. Nucleic Acids Res. 1975 Jun;2(6):915–930. doi: 10.1093/nar/2.6.915. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Donis-Keller H., Maxam A. M., Gilbert W. Mapping adenines, guanines, and pyrimidines in RNA. Nucleic Acids Res. 1977 Aug;4(8):2527–2538. doi: 10.1093/nar/4.8.2527. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dube S. K., Marcker K. A. The nucleotide sequence of N-formyl-methionyl-transfer RNA. Partial digestion with pancreatic and T-1 ribonuclease and derivation of the total primary structure. Eur J Biochem. 1969 Mar;8(2):256–262. doi: 10.1111/j.1432-1033.1969.tb00522.x. [DOI] [PubMed] [Google Scholar]
  9. England T. E., Gumport R. I., Uhlenbeck O. C. Dinucleoside pyrophosphate are substrates for T4-induced RNA ligase. Proc Natl Acad Sci U S A. 1977 Nov;74(11):4839–4842. doi: 10.1073/pnas.74.11.4839. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. England T. E., Uhlenbeck O. C. Enzymatic oligoribonucleotide synthesis with T4 RNA ligase. Biochemistry. 1978 May 30;17(11):2069–2076. doi: 10.1021/bi00604a008. [DOI] [PubMed] [Google Scholar]
  11. Gillam I., Blew D., Warrington R. C., von Tigerstrom M., Tener G. M. A general procedure for the isolation of specific transfer ribonucleic acids. Biochemistry. 1968 Oct;7(10):3459–3468. doi: 10.1021/bi00850a022. [DOI] [PubMed] [Google Scholar]
  12. Greenfield J. C., Leonard N. J., Gumport R. I. Nicotinamide 3,N4-ethenocytosine dinucleotide, an analog of nicotinamide adenine dinucleotide. Synthesis and enzyme studies. Biochemistry. 1975 Feb 25;14(4):698–706. doi: 10.1021/bi00675a009. [DOI] [PubMed] [Google Scholar]
  13. Kaufmann G., Littauer U. Z. Covalent joining of phenylalanine transfer ribonucleic acid half-molecules by T4 RNA ligase. Proc Natl Acad Sci U S A. 1974 Sep;71(9):3741–3745. doi: 10.1073/pnas.71.9.3741. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Lillehaug J. R., Kleppe K. Phosphorylation of tRNA by T4 polynucleotide kinase. Nucleic Acids Res. 1977 Feb;4(2):373–380. doi: 10.1093/nar/4.2.373. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Linné T., Oberg B., Philipson L. RNA ligase activity in phage-infected bacteria and animal cells. Eur J Biochem. 1974 Feb 15;42(1):157–165. doi: 10.1111/j.1432-1033.1974.tb03325.x. [DOI] [PubMed] [Google Scholar]
  16. Morris R. W., Herbert E. Purification and characterization of yeast nucleotidyl transferase and investigation of enzyme-transfer ribonucleic acid complex formation. Biochemistry. 1970 Nov 24;9(24):4819–4827. doi: 10.1021/bi00826a029. [DOI] [PubMed] [Google Scholar]
  17. Ohtsuka E., Nishikawa S., Sugiura M., Ikehara M. Joining of ribooligonucleotides with T4 RNA ligase and identification of the oligonucleotide-adenylate intermediate. Nucleic Acids Res. 1976 Jun;3(6):1613–1623. doi: 10.1093/nar/3.6.1613. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Silber R., Malathi V. G., Hurwitz J. Purification and properties of bacteriophage T4-induced RNA ligase. Proc Natl Acad Sci U S A. 1972 Oct;69(10):3009–3013. doi: 10.1073/pnas.69.10.3009. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Simoncsits A., Brownlee G. G., Brown R. S., Rubin J. R., Guilley H. New rapid gel sequencing method for RNA. Nature. 1977 Oct 27;269(5631):833–836. doi: 10.1038/269833a0. [DOI] [PubMed] [Google Scholar]
  20. Sninsky J. J., Last J. A., Gilham P. T. The use of terminal blocking groups for the specific joining of oligonucleotides in RNA ligase reactions containing equimolar concentrations of acceptor and donor molecules. Nucleic Acids Res. 1976 Nov;3(11):3157–3166. doi: 10.1093/nar/3.11.3157. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Uhlenbeck O. C., Cameron V. Equimolar addition of oligoribonucleotides with T4 RNA ligase. Nucleic Acids Res. 1977 Jan;4(1):85–98. doi: 10.1093/nar/4.1.85. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Walker G. C., Uhlenbeck O. C., Bedows E., Gumport R. I. T4-induced RNA ligase joins single-stranded oligoribonucleotides. Proc Natl Acad Sci U S A. 1975 Jan;72(1):122–126. doi: 10.1073/pnas.72.1.122. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Wimmer E., Maxwell I. H., Tener G. M. A simple method for isolating highly purified yeast phenylalanine transfer ribonucleic acid. Biochemistry. 1968 Jul;7(7):2623–2628. doi: 10.1021/bi00847a026. [DOI] [PubMed] [Google Scholar]
  24. Zachau H. G., Hertz H. S. Lack of incorporation of 1:N6-ethenoadenosine triphosphate into yeast tRNAPhe and tRNASer under standard conditions. Eur J Biochem. 1974 May 2;44(1):289–291. doi: 10.1111/j.1432-1033.1974.tb03484.x. [DOI] [PubMed] [Google Scholar]

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

RESOURCES