Nonenzymatic sequence-specific methyl transfer to single-stranded DNA.
2'-Deoxyuridine 5'-triphosphate with a methylthioether moiety at the 5 position can be incorporated into a primer-template DNA complex with Klenow enzyme. Activation with CNBr at 25 degrees C, pH 5.5, followed by treatment with piperidine produces sequence-specific cleavage on the template DNA predominantly at a single guanine position. The mechanism involves methyl-group transfer from sulfur on the modified deoxyuridine of the extended primer to N-7 of guanine on the template DNA. This raises the possibility for the design and synthesis of a nonenzymatic class of sequence-specific methyltransferases for DNA.
Iverson
B L
BL
Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena 91125.
Dervan
P B
PB
D001483:Base Sequence; D004277:DNA, Single-Stranded / Q000378:metabolism*; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism; D003856:Deoxyuracil Nucleotides / Q000138:chemical synthesis / Q000378:metabolism*; D006147:Guanine / Q000031:analogs & derivatives / Q000378:metabolism; D008745:Methylation; D008969:Molecular Sequence Data
D004277:DNA, Single-Stranded; D003856:Deoxyuracil Nucleotides; C056479:5-(3-((3-(methylthio)propionyl)amino)-1-propenyl)-2'-deoxyuridine-5'-triphosphate; D006147:Guanine; C008450:7-methylguanine; D004259:DNA-Directed DNA Polymerase