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Nucleic Acids Research, 1985, Vol. 13, No. 10 3479-3494
© 1985

Articles

DNA methylation: sequences flanking C-G pairs modulate the specificity of the human DNA methylase

Arthur H. Bolden, Carlo M. Nalin, Cheryl A. Ward, Mohinder S. Poonian+, Warren W. McComas+ and Arthur Weissbach*

Department of Cell Biology, Roche Institute of Molecular Biology, Roche Research Center Nutley, NJ 07110, USA +Department of Molecular Genetics, Hoffmann-La Roche Inc. Nutley, NJ 07110, USA

*To whom correspondaence should be addressed

Received February 19, 1985. Revised April 16, 1985. Accepted April 16, 1985.

Synthetic single-stranded oligodeoxynucleotides of known sequence have been used as in vitro substrates for a partially purified HeLa cell DNA methylase. Although most oligonucleotides tested cannot be used by the HeLa DNA methylase in vitro, we have found a unique 27mer, containing 2 C-G pairs, that is an excellent substrate for the enzyme. Analysis of the methylation of the 27mer, its derivatives and other oligomer substrates reveal that the HeLa DNA methylase does not significantly methylate an oligomer which contains just one C-G pair. In addition, only one of the two C-G pairs in the 27mer is methylated and this methylatlon is abolished if the other C-G pair is converted to a C-A pair. Furthermore, the HeLa enzyme apparently cannot methylate C-G pairs located in compounds containing a high A + T content. The most efficient methylatlon occurs with multiple separated C-G pairs in a compound with a high G + C content (>65%). The results suggest that clustering of C-G pairs in regions of the DNA high In G + C content may be the preferred site for DNA methylation in viva.


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