Use of EDTA Derivatization To Characterize Interactions between Oligodeoxyribonucleoside Methylphosphonates and Nucleic Acids

Shwu Bin Lin, Kathleen R. Blake, Paul S. Miller, Paul O.P. Ts'o

Research output: Contribution to journalArticlepeer-review

39 Scopus citations


EDTA-derivatized oligonucleoside methylphosphonates were prepared and used to characterize hybridization between the oligomers and single-stranded DNA or RNA. The melting temperatures of duplexes formed between an oligodeoxyribonucleotide 35-mer and complementary methylphosphonate 12-mers were 4-12 °C higher than those of duplexes formed by oligodeoxyribonucleotide 12-mers as determined by spectrophotometric measurements. Derivatization of the methylphosphonate oligomers with EDTA reduced the melting temperature by 5 °C. Methylphosphonate oligomer-nucleic acid complexes were stabilized by base stacking interactions between the terminal bases of the two oligomers binding to adjacent binding sites on the target. In the presence of Fe2+ and DTT, the EDTA-derivatized oligomers produce hydroxyl radicals that cause degradation of the sugar-phosphate backbone of both targeted DNA and RNA. Degradation occurs specifically in the region of the oligomer binding site and is approximately 20-fold more efficient for single-stranded DNA than for RNA. In comparison to the presence of one oligomer, the extent of target degradation was increased considerably by the additions of two oligomers that bind at adjacent sites on the target. For example, the extent or degradation of a single-stranded DNA 35-mer caused by two contiguously binding oligomers, one of which was derivatized by EDTA, was approximately 2 times greater than that causea by the EDTA-derivatized oligomer alone. Although EDTA-derivatized oligomers are stable for long periods of time in aqueous solution, they undergo rapid autodegradation in the presence of Fe2+ and DTT with half-lives of approximately 30 min. This autodegradation reaction renders the EDTA-derivatized oligomers unable to cause degradation of their complementary target nucleic acids. It appears that cleavage of the EDTA portion of the molecule by hydroxyl radicals is the major cause of this autodegradation and that the methylphosphonate backbone is resistant to cleavage.

Original languageEnglish (US)
Pages (from-to)1054-1061
Number of pages8
Issue number3
StatePublished - 1989
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry


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