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Nucleic Acids Research, 1982, Vol. 10, No. 15 4641-4653
© 1982

MOLECULAR BIOLOGY

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

David R. Sargan and Peter H.W. Butterworth

Department of Biochemistry, University College London Gower Street, London WC1E 6BT, UK

Received March 12, 1982. Revised June 7, 1982. Accepted July 5, 1982.

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 llS mono-nucleosome and the other with the dinucleosome. The released material can act as a source of ternary transcription complexes for further structural studies.


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