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Autodigestion of rat liver nuclei in magnesium-containing buffers leads to the release of about 80% of DNA-dependent RNA polymerases I and II, together with 4 to 8% of the DNA. The RNA polymerases are at least partially DNA bound as judged by the effect on in vitro transcription of (1) Actinomycin D, (2) preirradiation of the enzymes in the presence of 8-methoxypsoralen and (3) heparin. The released DNA migrates as ladders of nicked nucleosome-size fragments on electrophoresis. On sucrose gradients, most RNA polymerase activity sediments as two peaks: one slightly smaller than the 11S mononucleosome and the other with the dinucleosome. The released material can act as a source of ternary transcription complexes for further structural studies.

nucleic acids research

Eukaryotic ternary transcription complexes. I. The release of ternary transcription complexes of RNA polymerases I and II by the endogenous nucleases of rat liver nuclei