Human placental DNA methyltransferase: DNA substrate and DNA binding specificity.
We have partially purified a DNA methyltransferase from human placenta using a novel substrate for a highly sensitive assay of methylation of hemimethylated DNA. This substrate was prepared by extensive nick translation of bacteriophage XP12 DNA, which normally has virtually all of its cytosine residues replaced by 5-methylcytosine (m5C). Micrococcus luteus DNA was just as good a substrate if it was first similarly nick translated with m5dCTP instead of dCTP in the polymerization mixture. At different stages in purification and under various conditions (including in the presence or absence of high mobility group proteins), the methylation of m5C-deficient DNA and that of hemimethylated DNA were compared. Although hemimethylated , m5C-rich DNAs were much better substrates than were m5C-deficient DNAs and normal XP12 DNA could not be methylated, all of these DNAs were bound equally well by the enzyme. In contrast, from the same placental extract, a DNA-binding protein of unknown function was isolated which binds to m5C-rich DNA in preference to the analogous m5C-poor DNA.
D004248:DNA (Cytosine-5-)-Methyltransferases / Q000302:isolation & purification / Q000378:metabolism*; D004269:DNA, Bacterial / Q000235:genetics; D004279:DNA, Viral / Q000235:genetics; D004268:DNA-Binding Proteins / Q000302:isolation & purification / Q000378:metabolism; D005260:Female; D006801:Humans; D007700:Kinetics; D008780:Methyltransferases / Q000378:metabolism*; D008837:Micrococcus / Q000235:genetics; D010920:Placenta / Q000201:enzymology*; D011247:Pregnancy; D014176:Protein Biosynthesis; D013379:Substrate Specificity
D004269:DNA, Bacterial; D004279:DNA, Viral; D004268:DNA-Binding Proteins; D008780:Methyltransferases; D004248:DNA (Cytosine-5-)-Methyltransferases