Characterization of the DNA methylase activity of the restriction enzyme from Escherichia coli K.
The restriction endonuclease from Escherichia coli K is a multifunctional protein which efficiently methylates heteroduplex DNA (one strand modified and one strand unmodified) in the presence of S-adenosylmethionine (AdoMet), ATP, and Mg2+. The methylase activity is catalytic, and seems to modify different heteroduplex host specificity sites for E. coli K with equal efficiency. In the methylase reaction, both AdoMet and ATP (or its imido analog) act as allosteric effectors, but AdoMet also serves as a methyl donor. Preincubation of the enzyme with AdoMet eliminates the lag period observed in DNA methylation. The rate of enzyme activation was determined using the AdoMet analog Sinefungin. The result are consistent with the hypothesis that the early steps of AdoMet binding and enzyme activation are common to both restriction and modification reactions.
D010582:Bacteriophage lambda; D004248:DNA (Cytosine-5-)-Methyltransferases / Q000378:metabolism*; D004262:DNA Restriction Enzymes / Q000378:metabolism*; D004279:DNA, Viral; D015253:Deoxyribonucleases, Type I Site-Specific; D004789:Enzyme Activation; D004926:Escherichia coli / Q000201:enzymology*; D007700:Kinetics; D008780:Methyltransferases / Q000378:metabolism*; D012436:S-Adenosylmethionine / Q000494:pharmacology
D004279:DNA, Viral; D012436:S-Adenosylmethionine; D008780:Methyltransferases; D004248:DNA (Cytosine-5-)-Methyltransferases; D004262:DNA Restriction Enzymes; C029602:endodeoxyribonuclease EcoK; D015253:Deoxyribonucleases, Type I Site-Specific