Determination of methylation site of DNA-methyltransferase NlaX by a hybrid method.
Using a new method based on a combination of bisulfite reaction, the repair enzyme uracil-DNA glycosylase, and synthetic oligodeoxyribonucleotides, the methylation site of DNA-methyltransferase NlaX (M.NlaX) from Neisseria lactamica was established to be the inner cytosine in the double-stranded pentanucleotide recognition sequence 5'-CCNGG-3' (where N = any nucleoside). 5-Methylcytosine (m5C) type modification by M-N1aX was confirmed by the use of oligonucleotide substrates that contain 5-fluoro-2'-deoxycytidine.
Kubareva
E A
EA
Institute of Physico-Chemical Biology, Moscow State University, Russia. kubareva@belozersky.msu.ru
Walter
J
J
Karyagina
A S
AS
Vorob'eva
O V
OV
Lau
P C K
PC
Trautner
T
T
D002623:Chemistry Techniques, Analytical / Q000379:methods*; D003432:Cross-Linking Reagents / Q000737:chemistry; D045647:DNA Glycosylases; D019175:DNA Methylation; D015254:DNA Modification Methylases / Q000737:chemistry* / Q000302:isolation & purification; D004269:DNA, Bacterial / Q000737:chemistry; D015252:Deoxyribonucleases, Type II Site-Specific / Q000737:chemistry; D009699:N-Glycosyl Hydrolases / Q000737:chemistry*; D009343:Neisseria / Q000737:chemistry / Q000201:enzymology / Q000235:genetics; D013447:Sulfites / Q000737:chemistry; D051981:Uracil-DNA Glycosidase
D003432:Cross-Linking Reagents; D004269:DNA, Bacterial; D013447:Sulfites; D015254:DNA Modification Methylases; C048422:endodeoxyribonuclease NlaIII; C048423:endodeoxyribonuclease NlaIV; D015252:Deoxyribonucleases, Type II Site-Specific; D045647:DNA Glycosylases; D009699:N-Glycosyl Hydrolases; D051981:Uracil-DNA Glycosidase