Stereochemical course of DNA hydrolysis by nuclease S1

Nuclease S1 hydrolyzes the Sp-diastereomer of 5'-O-(2'-deoxyadenosyl)-3'-O-thymidyl phosphorothioate in H2(18)O to [18O]deoxyadenosine 5'-O-phosphorothioate which can be phosphorylated enzymatically to the Sp-diastereomer of [alpha-18O]deoxyadenosine 5'-O-(1-thiotriphosphate...

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Bibliographic Details
Published in:The Journal of biological chemistry 1983-02, Vol.258 (3), p.1758-1760
Main Authors: Potter, B V, Romaniuk, P J, Eckstein, F
Format: Article
Language:English
Subjects:
Aspergillus oryzae - enzymology
Chromatography, High Pressure Liquid
Endonucleases - metabolism
Kinetics
Oligodeoxyribonucleotides
Oligonucleotides
Oxygen Isotopes
Single-Strand Specific DNA and RNA Endonucleases
Substrate Specificity
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Summary:Nuclease S1 hydrolyzes the Sp-diastereomer of 5'-O-(2'-deoxyadenosyl)-3'-O-thymidyl phosphorothioate in H2(18)O to [18O]deoxyadenosine 5'-O-phosphorothioate which can be phosphorylated enzymatically to the Sp-diastereomer of [alpha-18O]deoxyadenosine 5'-O-(1-thiotriphosphate). 31P nmr spectroscopy shows the oxygen-18 in this compound to be in a nonbridging position at the alpha-phosphorus, indicating that the hydrolysis reaction catalyzed by nuclease S1 proceeds with inversion of configuration at phosphorus. This result is compatible with a direct nucleophilic attack of H2O at phosphorus without the involvement of a covalent enzyme intermediate.
ISSN:0021-9258
1083-351X
DOI:10.1016/S0021-9258(18)33051-5