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Phosphorothioation of Oligonucleotides Strongly Influences the Inhibition of Bacterial (M.HhaI) and Human (Dnmt1) DNA Methyltransferases
Author(s) -
Warncke Simon,
Gégout Aline,
Carell Thomas
Publication year - 2009
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.200800798
Subject(s) - methyltransferase , dnmt1 , dna methyltransferase , dna , oligonucleotide , cpg site , phosphodiester bond , dna methylation , cytidine , biochemistry , chemistry , endonuclease , methylation , biology , microbiology and biotechnology , enzyme , gene , rna , gene expression
Methyltransferase inhibitors : Short double‐stranded oligonucleotides that have a hemimethylated target sequence and 5‐fluoro‐2′‐deoxycytidine as a suicide inhibitor as well as their phosphorothioated analogues were tested for their ability to inhibit the bacterial methyltransferase M.HhaI and the human Dnmt1 in vitro.The cytidine analogue 5‐fluoro‐2′‐deoxycytidine (dC F ) is a mechanism‐based inhibitor of DNA methyltransferases. We report the synthesis of short 18‐mer dsDNA oligomers containing a triple‐hemimethylated CpG motive as a recognition sequence for the human methyltransferase Dnmt1. The DNA strands carry within these CpG islands dC F building blocks that function as mechanism‐based inhibitors of the analyzed methyltransferases. In addition, we replaced the phosphodiester backbones at defined positions by phosphorothioates. These hypermodified DNA strands were investigated as inhibitors of the DNA methyltransferases M.HhaI and Dnmt1 in vitro. We could show that both methylases behave substantially differently in respect to the amount of DNA backbone modification.