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Biochemistry ; 39(16): 4939-46, 2000 Apr 25.
Article in English | MEDLINE | ID: mdl-10769153

ABSTRACT

Solid-state deuterium NMR is used to investigate perturbations of the local, internal dynamics in the EcoRI restriction binding site, -GAATTC- induced by cytidine methylation. Methylation of the cytidine base in this sequence is known to suppress hydrolysis by the EcoRI restriction enzyme. Previous solid-state deuterium NMR studies have detected large amplitude motions of the phosphate-sugar backbone at the AT-CG junction of the unmethylated DNA sequence. This study shows that methylation of the cytidine base in a CpG dinucleotide reduces the amplitudes of motions of the phosphate-sugar backbone. These observations suggest a direct link between suppression of the amplitudes of localized, internal motions of the sugar-phosphate backbone of the DNA and inhibition of restriction enzyme cleavage.


Subject(s)
CpG Islands/genetics , DNA Methylation , DNA/chemistry , DNA/metabolism , Base Sequence , Computer Simulation , Cytidine/chemistry , Cytidine/genetics , Cytidine/metabolism , DNA/genetics , Deoxyribonuclease EcoRI/metabolism , Deuterium/metabolism , Diffusion , Kinetics , Magnetic Resonance Spectroscopy , Motion , Structure-Activity Relationship , Substrate Specificity
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