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J. Biol. Chem., Vol. 259, Issue 20, 12437-12443, 10, 1984
A Bolden, C Ward, JA Siedlecki and A Weissbach
A partially purified HeLa cell DNA methylase will methylate a totally
unmethylated DNA (de novo methylation) at about 3-4% the rate it will
methylate a hemimethylated DNA template (maintenance methylation). Our
evidence suggests that many, if not most, dCpdG sequences in a natural or
synthetic DNA can be methylated by the enzyme. There is a powerful
inhibitor of DNA methylase activity in crude extracts which has been
identified as RNA. The inhibition of DNA methylase by RNA may indicate that
this enzyme is regulated in vivo by the presence of RNA at specific
chromosomal sites. The pattern of binding of RNA to DNA in the nucleosome
structure and the DNA replication complex may determine specific sites of
DNA methylation. An even more potent inhibition of DNA methylase activity
is observed with poly(G), but not poly(C), poly(A), or poly(U). The only
other synthetic polynucleotides studied which inhibit DNA methylation as
well as poly(G) are the homopolymers poly(dC).poly(dG) and poly
(dA).poly(dT). These results point out the unique importance of the guanine
residue itself in the binding of the DNA methylase to dCpdG, the site of
cytosine methylation. The surprising inhibition of the methylation reaction
by poly(dA).poly(dT), which is itself not methylated by the enzyme,
suggests the possible involvement of adjacent A and T residues in
influencing the choice of sites of methylation by the enzyme.
DNA methylation. Inhibition of de novo and maintenance methylation in vitro by RNA and synthetic polynucleotides
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