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Nucleic Acids Research, 1984, Vol. 12, No. 21 8161-8180
© 1984

MOLECULAR BIOLOGY

In vitro evidence that eukaryotic ribosomal RNA transcription is regulated by modification of RNA polymerase I

Marvin R. Paule, Calvin T. Iida, Peter J. Perna, Guy H. Harris, Deborah A. Knoll and J.M. D'Alessio

Department of Biochemistry and Cellular and Molecular Biology Interdisciplinary Program, Colorado State University Fort Collins, CO 80523, USA

Received February 17, 1984. Revised October 3, 1984. Accepted October 3, 1984.

We have utilized a cell-free transcription system from Acanthamoeba castellanii to test the functional activity of RNA polymerase I and transcription initiation factor I (TIF-I) during developmental down regulation of rRNA transcription. The results strongly suggest that rRKA transcription is regulated by modification, probably covalent, of RKA polymerase I: (1) The level of activity of TIF-I in extracts from transcriptionally active and inactive cells is constant. (2) The number of RNA polymerase I molecules in transcriptionally active and inactive cells is also constant. (3) In contrast, though the specific activity of polymerase I on damaged templates remains constant, both crude and purified polymerase I from inactive cells have lost the ability to participate in faithful initiation of rRNA transcription. (4) Polymerase I purified from transcriptionally active cells has the same subunit architecture as enzyme from inactive cells. However, the latter is heat denatured 5 times faster than the active polynerase.


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