Effects of 5-(N-aminohexyl)carbamoyl-2'-deoxyuridine on endonuclease stability and the ability of oligodeoxynucleotide to activate RNase H

Nucleic Acids Res. 1997 Oct 1;25(19):3777-82. doi: 10.1093/nar/25.19.3777.

Abstract

To evaluate an endonuclease resistance property of oligodeoxynucleotides (ODNs) containing 5-(N-aminohexyl)carbamoyl-2'-deoxyuridines (Hs) and to elucidate whether a duplex consisting of the ODN analogue and its complementary RNA induces RNase H activity, the ODNs containing the deoxyuridine analogues, Hs, at intervals of one, two, three, four and five natural nucleosides were synthesized. From partial hydrolysis of these ODNs with nuclease S1 (an endonuclease), it was found that the ODNs became more stable towards nucleolytic hydrolysis by the enzyme as the number of H increased. Furthermore, to examine whether the duplexes composed of the ODNs containing Hs and their complementary RNAs are substrates for RNase H or not, the duplexes of these ODNs and their complementary RNA strands were treated with Escherichia coliRNase H. It was found that cleavage of the RNA strands by the enzyme was kinetically affected by the introduction of Hs into the duplexes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Circular Dichroism
  • Deoxyuridine / analogs & derivatives*
  • Deoxyuridine / pharmacology
  • Electrophoresis, Polyacrylamide Gel
  • Enzyme Activation / drug effects
  • Enzyme Stability / drug effects
  • Escherichia coli / enzymology
  • Hydrolysis
  • Kinetics
  • Magnetic Resonance Spectroscopy
  • Oligonucleotides, Antisense / genetics
  • Oligonucleotides, Antisense / pharmacology*
  • RNA, Complementary / metabolism
  • Ribonuclease H / metabolism*
  • Single-Strand Specific DNA and RNA Endonucleases / metabolism*
  • Spectrometry, Mass, Fast Atom Bombardment
  • Substrate Specificity
  • Thermodynamics

Substances

  • 5-(N-aminohexyl)carbamoyl-2'-deoxyuridine
  • Oligonucleotides, Antisense
  • RNA, Complementary
  • Ribonuclease H
  • Single-Strand Specific DNA and RNA Endonucleases
  • Deoxyuridine