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Nucleic Acids Res. 2002 Sep 1;30(17):3880-5.

Promiscuous methylation of non-canonical DNA sites by HaeIII methyltransferase.

Author information

1
MRC Centre for Protein Engineering and MRC Laboratory for Molecular Biology, MRC Centre, Hills Road, Cambridge CB2 2QH, UK and. Department of Biological Chemistry, The Weizmann Institute of Science, Rehovot 76 100, Israel.

Abstract

The cytosine C5 methyltransferase M.HaeIII recognises and methylates the central cytosine of its canonical site GGCC. Here we report that M.HaeIII can also, with lower efficiency, methylate cytosines located in a wide range of non-canonical sequences. Using bisulphite sequencing we mapped the methyl- cytosine residues in DNA methylated in vitro and in vivo by M.HaeIII. Methyl-cytosine residues were observed in multiple sequence contexts, most commonly, but not exclusively, at star sites (sites differing by a single base from the canonical sequence). The most frequently used star sites had changes at positions 1 and 4, but there is little or no methylation at star sites changed at position 2. The rate of methylation of non-canonical sites can be quite significant: a DNA substrate lacking a canonical site was methylated by M.HaeIII in vitro at a rate only an order of magnitude slower than an otherwise identical substrate containing the canonical site. In vivo methylation of non-canonical sites may therefore be significant and may have provided the starting point for the evolution of restriction-modification systems with novel sequence specificities.

PMID:
12202773
PMCID:
PMC137429
DOI:
10.1093/nar/gkf507
[Indexed for MEDLINE]
Free PMC Article

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