MULTIPLE NUCLEOSOME POSITIONING WITH UNIQUE ROTATIONAL SETTING FOR THE SACCHAROMYCES-CEREVISIAE 5S RIBOSOMAL-RNA GENE IN-VITRO AND IN-VIVO

A simple no-background assay was developed for high-resolution in vivo analysis of yeast chromatin. When applied to Saccharomyces cerevisiae 5S rRNA genes (5S rDNA), this analysis shows that nucleosomes completely cover this chromosomal region, occupying alternative positions characterized by a unique helical phase. This supports the notion that sequence-intrinsic rotational signals are the major determinant of nucleosome localization. Nucleosomal core particles reconstituted in vitro occupy the same positions and have the same helically phase distribution observed in vivo, as determined by mapping of exonuclease III-resistant borders, mapping by restriction cleavages, and by DNase I and hydroxyl-radical digestion patterns.