Structure of the active nucleolar chromatin of Xenopus laevis Oocytes.

Active nucleolar chromatin of Xenopus laevis oocytes was prepared for electron microscopy by a step gradient method, which separates the chromatin from proteins and other constituents that might nonspecifically bind at low ionic strength. Between putative RNA polymerases and within the nontranscribed spacer region, the chromatin appears as smooth, thin filaments. For the first time, it is shown here that these filaments are indistinguishable from pure DNA absorbed to the same specimen, even when the ionic strength is raised up to 100 mM NaCl. Bulk rat liver chromatin, however, which was coprepared as a biochemically well-characterized standard with the active nucleolar chromatin, shows nucleosomes containing fibers, which condense into supranucleosomal structures with increasing ionic strength. Since the appearance and the behavior of active nucleolar chromatin at different ionic strengths and pHs resembles tht of pure DNA, but not of any known type of chromatin, it is suggested that, except for the transcription apparatus, very few macromolecular constituents are associated with ribosomal DNA during transcription. The observations described in this paper explain most of the published and partly conflicting results obtained by electron microscopy of nucleolar chromatin.