Conversion of DNA methyltransferases into azidonucleosidyl transferases via synthetic cofactors.
Aziridine-based cofactor mimics have been synthesized and are shown to undergo methyltransferase-dependent DNA alkylation. Notably, each cofactor mimic possesses an azide functionality, to which can be attached an assortment of unnatural groups following methyltransferase-dependent DNA delivery. DNA duplexes modified with these cofactor mimics are capable of undergoing the Staudinger ligation with phosphines tethered to biological functionalities following enzymatic modification. This methodology provides a new tool by which to selectively modify DNA in a methyltransferase-dependent way. The conversion of biological methyltransferases into azidonucleosidyl transferases demonstrated here also holds tremendous promise as a means of identifying, as yet, unknown substrates of methylation.
Comstock
Lindsay R
LR
School of Pharmacy, University of Wisconsin-Madison, 777 Highland Ave., Madison, WI 53705, USA.
Rajski
Scott R
SR
D000478:Alkylation; D001386:Azides / Q000737:chemistry*; D001388:Aziridines / Q000737:chemistry*; D019921:Biotinylation; D003067:Coenzymes / Q000737:chemistry*; D004247:DNA / Q000737:chemistry / Q000378:metabolism*; D018716:Molecular Mimicry; D009705:Nucleosides / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism*; D009841:Oligonucleotides / Q000737:chemistry / Q000378:metabolism; D010430:Pentosyltransferases / Q000378:metabolism*; D015265:Site-Specific DNA-Methyltransferase (Adenine-Specific) / Q000378:metabolism*
D001386:Azides; D001388:Aziridines; D003067:Coenzymes; D009705:Nucleosides; D009841:Oligonucleotides; D004247:DNA; C020098:DNA modification methylase EcoRI; C059680:DNA modification methylase TaqI; D015265:Site-Specific DNA-Methyltransferase (Adenine-Specific); D010430:Pentosyltransferases