Affinity modification of EcoRII DNA methyltransferase by the dialdehyde-substituted DNA duplexes: mapping the enzyme region that interacts with DNA.
Affinity modification of EcoRII DNA methyltransferase (M x EcoRII) by DNA duplexes containing oxidized 2'-O-beta-D-ribofuranosylcytidine (Crib*) or 1-(beta-D-galactopyranosyl)thymine (Tgal*) residues was performed. Cross-linking yields do not change irrespective of whether active Crib* replaces an outer or an inner (target) deoxycytidine within the EcoRII recognition site. Chemical hydrolysis of M x EcoRII in the covalent cross-linked complex with the Tgal*-substituted DNA indicates the region Gly268-Met391 of the methylase that is likely to interact with the DNA sugar-phosphate backbone. Both specific and non-specific DNA interact with the same M x EcoRII region. Our results support the theoretically predicted DNA binding region of M x EcoRII.
Gritsenko
Oksana M
OM
Department of Chemistry, Moscow State University, Moscow, 119992, Russia.
Koudan
Elizaveta V
EV
Mikhailov
Sergey N
SN
Ermolinsky
Boris S
BS
Van Aerschot
Arthur
A
Herdewijn
Piet
P
Gromova
Elizaveta S
ES
D000447:Aldehydes / Q000378:metabolism*; D001483:Base Sequence; D001665:Binding Sites; D003432:Cross-Linking Reagents / Q000737:chemistry / Q000378:metabolism; D004247:DNA / Q000737:chemistry / Q000378:metabolism*; D019175:DNA Methylation; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000378:metabolism*; D015257:DNA-Cytosine Methylases / Q000737:chemistry* / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000737:chemistry / Q000378:metabolism
D000447:Aldehydes; D003432:Cross-Linking Reagents; D004268:DNA-Binding Proteins; D009838:Oligodeoxyribonucleotides; D004247:DNA; C029528:DNA modification methylase EcoRII; D015257:DNA-Cytosine Methylases