Solid phase synthesis and restriction endonuclease cleavage of oligodeoxynucleotides containing 5-(hydroxymethyl)-cytosine

Séverine Tardy-Planechaud, June Fujimoto, Susan S. Lin, Lawrence C. Sowers

Research output: Contribution to journalArticle

70 Scopus citations

Abstract

Emerging data suggest an important role for cytosine methylation in tumorigenesis. Simultaneously, recent studies indicate a significant contribution of endogenous oxidative DNA damage to the development of human disease. Oxidation of the 5-methyl group of 5-methylcytosine (5mC) residues in DNA results in the formation of 5-(hydroxymethyl)cytosine ((hm)C). The biological consequences of (hm)C residues in vertebrate DNA are as yet unknown; however, conversion of the hydrophobic methyl group to the hydrophilic hydroxymethyl group may substantially alter the interaction of sequence-specific binding proteins with DNA. Central to both biophysical and biochemical studies on the potential consequences of specific DNA damage products such as (hm)C are efficient methods for the synthesis of oligodeoxynucleotides containing such modified bases at selected positions. In this paper, we describe a method for the placement of (hm)C residues in oligodeoxynucleotides using established phosphoramidite chemistry. In addition, we have examined the influence of specific (hm)C residues on enzymatic cleavage of oligodeoxynucleotides by the methylation-sensitive restriction endonucleases MspI and HpaII.

Original languageEnglish (US)
Pages (from-to)553-558
Number of pages6
JournalNucleic acids research
Volume25
Issue number3
DOIs
StatePublished - 1997

ASJC Scopus subject areas

  • Genetics

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