M.HhaI binds tightly to substrates containing mismatches at the target base.

Abstract:

:The (cytosine-5) DNA methyltransferase M.HhaI causes its target cytosine base to be flipped completely out of the DNA helix upon binding. We have investigated the effects of replacing the target cytosine by other, mismatched bases, including adenine, guanine, thymine and uracil. We find that M.HhaI binds more tightly to such mismatched substrates and can even transfer a methyl group to uracil if a G:U mismatch is present. Other mismatched substrates in which the orphan guanine is changed exhibit similar behavior. Overall, the affinity of DNA binding correlates inversely with the stability of the target base pair, while the nature of the target base appears irrelevant for complex formation. The presence of a cofactor analog. S-adenosyl-L-homocysteine, greatly enhances the selectivity of the methyltransferase for cytosine at the target site. We propose that the DNA methyltransferases have evolved from mismatch binding proteins and that base flipping was, and still is, a key element in many DNA-enzyme interactions.

journal_name

Nucleic Acids Res

journal_title

Nucleic acids research

authors

Klimasauskas S,Roberts RJ

doi

10.1093/nar/23.8.1388

subject

Has Abstract

pub_date

1995-04-25 00:00:00

pages

1388-95

issue

8

eissn

0305-1048

issn

1362-4962

pii

4j0812

journal_volume

23

pub_type

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