Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1975 Jan;72(1):122–126. doi: 10.1073/pnas.72.1.122

T4-induced RNA ligase joins single-stranded oligoribonucleotides.

G C Walker, O C Uhlenbeck, E Bedows, R I Gumport
PMCID: PMC432253  PMID: 1090929

Abstract

RNA ligase isolated from Escherichia coli infected with bacteriophage T4 will catalyze the formation of an intermolecular 3' leads to 5' phosphodiester linkage between an oligoribonucleotide with a free 3'-hydroxyl and another oligoribonucleotide with a 5'-phosphate. Upon reaction with (Ap)5C, nearly quantitative conversion of the hexamer [5'-32P]p(Up)5U to the dodecamer (Ap)5C[3' leads to 5'-32P]p(Up)5U was observed. The product was identified by its mobility on RPC-5 column chromatography, its resistance to alkaline phosphatase, and the appearance of the expected radiolabeled products on hydrolysis with alkali, ribonuclease A, snake venom phosphodiesterase, and spleen phosphodiesterase. The coupling of other pairs of single-stranded oligoribonucleotides has also been demonstrated. The intermolecular joining reaction is probably mechanistically similar to the intramolecular cyclization activity previously reported for Tr RNA ligase. It is expected that this enzyme will be useful for the synthesis of RNA fragments of defined sequence.

Full text

PDF
122

Selected References

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

  1. Baltz R. H., Drake J. W. Bacteriophage T4 transformation: an assay for mutations induced in vitro. Virology. 1972 Aug;49(2):462–474. doi: 10.1016/0042-6822(72)90498-9. [DOI] [PubMed] [Google Scholar]
  2. Borer P. N., Uhlenbeck O. C., Dengler B., Tinoco I., Jr Circular dichroism of double-helical oligoribonucleotides. J Mol Biol. 1973 Nov 15;80(4):759–771. doi: 10.1016/0022-2836(73)90208-8. [DOI] [PubMed] [Google Scholar]
  3. Brutlag D., Kornberg A. Enzymatic synthesis of deoxyribonucleic acid. 36. A proofreading function for the 3' leads to 5' exonuclease activity in deoxyribonucleic acid polymerases. J Biol Chem. 1972 Jan 10;247(1):241–248. [PubMed] [Google Scholar]
  4. Fareed G. C., Wilt E. M., Richardson C. C. Enzymatic breakage and joining of deoxyribonucleic acid. 8. Hybrids of ribo- and deoxyribonucleotide homopolymers as substrates for polynucleotide ligase of bacteriophage T4. J Biol Chem. 1971 Feb 25;246(4):925–932. [PubMed] [Google Scholar]
  5. Glynn I. M., Chappell J. B. A simple method for the preparation of 32-P-labelled adenosine triphosphate of high specific activity. Biochem J. 1964 Jan;90(1):147–149. doi: 10.1042/bj0900147. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Kaufmann G., Fridkin M., Zutra A., Littauer U. Z. Monofunctional substrates of polynucleotide phosphorylase. The monoaddition of 2'(3')-O-isovaleryl-nucleoside diphosphate to an initiator oligonucleotide. Eur J Biochem. 1971 Dec 22;24(1):4–11. doi: 10.1111/j.1432-1033.1971.tb19649.x. [DOI] [PubMed] [Google Scholar]
  7. Khorana H. G., Agarwal K. L., Büchi H., Caruthers M. H., Gupta N. K., Kleppe K., Kumar A., Otsuka E., RajBhandary U. L., Van de Sande J. H. Studies on polynucleotides. 103. Total synthesis of the structural gene for an alanine transfer ribonucleic acid from yeast. J Mol Biol. 1972 Dec 28;72(2):209–217. doi: 10.1016/0022-2836(72)90146-5. [DOI] [PubMed] [Google Scholar]
  8. Kirkegaard L. H. Gradient sievorptive chromatography. A focusing system for the separation of cellular components. Biochemistry. 1973 Sep 11;12(19):3627–3632. doi: 10.1021/bi00743a009. [DOI] [PubMed] [Google Scholar]
  9. LINN S., LEHMAN I. R. AN ENDONUCLEASE FROM NEUROSPORA CRASSA SPECIFIC FOR POLYNUCLEOTIDES LACKING AN ORDERED STRUCTURE. I. PURIFICATION AND PROPERTIES OF THE ENZYME. J Biol Chem. 1965 Mar;240:1287–1293. [PubMed] [Google Scholar]
  10. 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]
  11. Mackey J. K., Gilham P. T. New approach to the synthesis of polyribonucleotides of defined sequence. Nature. 1971 Oct 22;233(5321):551–553. doi: 10.1038/233551a0. [DOI] [PubMed] [Google Scholar]
  12. Modrich P., Lehman I. R. Enzymatic joining of polynucleotides. IX. A simple and rapid assay of polynucleotide joining (ligase) activity by measurement of circle formation from linear deoxyadenylate-deoxythymidylate copolymer. J Biol Chem. 1970 Jul 25;245(14):3626–3631. [PubMed] [Google Scholar]
  13. Olivera B. M., Lehman I. R. Enzymic joining of polynucleotides. 3. The polydeoxyadenylate-polydeoxythymidylate homopolymer pair. J Mol Biol. 1968 Sep 14;36(2):261–274. doi: 10.1016/0022-2836(68)90380-x. [DOI] [PubMed] [Google Scholar]
  14. Olivera B. M., Scheffler I. E., Lehman I. R. Enzymic joining of polynucleotides. IV. Formation of a circular deoxyadenylate-deoxythymidylate copolymer. J Mol Biol. 1968 Sep 14;36(2):275–285. doi: 10.1016/0022-2836(68)90381-1. [DOI] [PubMed] [Google Scholar]
  15. Pearson R. L., Weiss J. F., Kelmers A. D. Improved separation of transfer RNA's on polychlorotrifuoroethylene-supported reversed-phase chromatography columns. Biochim Biophys Acta. 1971 Feb 11;228(3):770–774. doi: 10.1016/0005-2787(71)90748-9. [DOI] [PubMed] [Google Scholar]
  16. Richardson C. C. Phosphorylation of nucleic acid by an enzyme from T4 bacteriophage-infected Escherichia coli. Proc Natl Acad Sci U S A. 1965 Jul;54(1):158–165. doi: 10.1073/pnas.54.1.158. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. 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]
  18. Silver M. J., Rodalewicz I., Duglas Y., Park D. High-voltage paper electrophoresis of nucleotides. Anal Biochem. 1970 Aug;36(2):525–527. doi: 10.1016/0003-2697(70)90389-1. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES