There are about 50 copies of each chromosome in the somatic macronucleus of the ciliated protozoan Tetrahymena. Approximately 0.8% of the adenine residues in the macronuclear DNA of Tetrahymena are methylated to N(6)-methyladenine. The degree of methylation varies between sites from a very low percentage to >90%. In this study a correlation was found between nucleosome positioning and DNA methylation. Eight GATC sites with different levels of methylation were examined. There was a direct correlation between the degree of methylation and proximity to linker DNA at these sites. Although methylation occurs preferentially in linker DNA, the patterns and extent of methylation in a histone H1 knockout strain were virtually indistinguishable from those in wild-type cells