S-adenosyl methionine alters the DNA contacts of the EcoKI methyltransferase.
The EcoKI methyltransferase methylates two adenines on opposite strands of its bipartite DNA recognition sequence AAC(N6)GTGC. The enzyme has a strong preference for hemimethylated DNA substrates, but the methylation state of the DNA does not influence its binding affinity. Methylation interference was used to compare the contacts made by the EcoKI methyltransferase with unmodified, hemimethylated or fully modified DNAs. Contacts were seen at or near the N7 position of guanine, in the major groove, for all of the guanines in the EcoKI recognition sequence, and at two guanines on the edge of the intervening spacer sequence. The presence of the cofactor and methyl donor S-adenosyl methionine had a striking effect on the interference pattern for unmodified DNA which could not be mimicked by the presence of the cofactor analogue S-adenosyl homocysteine. In contrast, S-adenosyl methionine had no effect on the interference patterns for either kind of hemimethylated DNA, or for fully modified DNA. Differences between the interference patterns for the unmodified DNA and any of the three forms of methylated DNA provide evidence that methylation of the target sequence influences the conformation of the protein-DNA interface, and illustrate the importance of S-adenosyl methionine in the distinction between unmodified and methylated DNA by the methyltransferase.
Powell
L M
LM
Institute of Cell and Molecular Biology, University of Edinburgh, UK.
Murray
N E
NE
D001483:Base Sequence; D004247:DNA / Q000737:chemistry / Q000378:metabolism*; D008745:Methylation; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D012435:S-Adenosylhomocysteine / Q000378:metabolism; D012436:S-Adenosylmethionine / Q000378:metabolism*; D015265:Site-Specific DNA-Methyltransferase (Adenine-Specific) / Q000378:metabolism*; D013463:Sulfuric Acid Esters; D013696:Temperature
D013463:Sulfuric Acid Esters; D012436:S-Adenosylmethionine; D004247:DNA; D012435:S-Adenosylhomocysteine; C056438:DNA modification methylase EcoK; D015265:Site-Specific DNA-Methyltransferase (Adenine-Specific); C007482:dimethyl sulfate