Methylation Of Chromatin Dna

E. coli DNA methylase has been used to methylate chromatin DNA in vitro. At saturation only 50% of the chromatin DNA becomes methylated. The methylated regions of chromatin correspond to that fraction of the chromatin which is sensitive to staphylococcal nuclease. Using in vitro methylated chromatin followed by nuclease digestion movement of chromatin proteins along the DNA can be detected. By this criterion, sonication of chromatin or precipitation with MnCl causes 10% of the previously uncovered methylated regions to become covered by protein. Reconstitution of methylated chromatin results in the randomization of the chromatin proteins. Using nuclei which were methylated in vitro we have demonstrated that a small degree of protein sliding does occur during the preparation of chromatin from nuclei. Finally, we have prepared open region DNA by polylysine titration. This procedure does not cause displacement of chromatin proteins.