Addition of mononucleotides to oligoribonucleotide acceptors with T4 RNA ligase.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 75 ·No. 3 ·1978-03-00 ·Pages 1270-3

Kikuchi Y, Hishinuma F, Sakaguchi K

Abstract

RNA ligase induced by bacteriophage T4 catalyzed the addition of nucleoside 5',3'-diphosphates to oligoribonucleotide acceptors in the presence of ATP. The reactions proceeded in equimolar concentrations of donors and acceptors. Several oligonucleotides of defined sequence were synthesized by this method in yields of 28-68%. The enzyme required the phosphate ester at the 3' position of the donor molecule, nucleoside 5',2'-diphosphates being unable to serve as donors. Thymidine 5',3'-diphosphate was active as a donor for the enzyme. The ligation product, (Ap)2ApCp, was also obtained from the reaction of (Ap)2A and 5'-adenylylated cytidine 5',3'-diphosphate (A5'pp5'Cp) with RNA ligase in the absence of ATP. These results show that the minimal substrate for RNA ligase is a nucleoside 5',3'-diphosphate.

MeSH Terms

Adenosine Triphosphate/metabolism Coliphages/enzymology Nucleotides/metabolism Oligonucleotides/metabolism Oligoribonucleotides/metabolism Polynucleotide Ligases/metabolism RNA Ligase (ATP)/metabolism Substrate Specificity

Chemicals

Nucleotides Oligonucleotides Oligoribonucleotides Adenosine Triphosphate Polynucleotide Ligases RNA Ligase (ATP)

Authors & Affiliations

3 authors, click to expand affiliations / ORCID

Kikuchi Y

Hishinuma F

Sakaguchi K

References (22)

22 references, click to expand

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Article Info

Journal

Proceedings of the National Academy of Sciences of the United States of America

Abbr.

Proc Natl Acad Sci U S A

ISSN

0027-8424

DOI

10.1073/pnas.75.3.1270

Published

1978-03-00

Pages

1270-3

Language

English

Region

United States

NLM ID

7505876

PMCID

PMC411452

Subset

External Links

PubMed source DOI: 10.1073/pnas.75.3.1270 PMC Full Text View journal details

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Addition of mononucleotides to oligoribonucleotide acceptors with T4 RNA ligase.

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