Specific transcription of mouse ribosomal DNA in a cell-free system that mimics control in vivo.
Open Access
- 1 February 1981
- journal article
- research article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences
- Vol. 78 (2), 727-731
- https://doi.org/10.1073/pnas.78.2.727
Abstract
Cloned rDNA from mouse, which contains the initiation site of 45S pre-rRNA transcription and 5'' flanking sequences, was used as the template in an in vitro transcription system. In the presence of extracts from rapidly growing Ehrlich ascites cells, RNA polymerase I initiates specifically in that region of purified rDNA where the 5'' end of 45S rRNA was mapped. This is shown by electrophoretic analysis of the length of run-off transcripts synthesized from truncated templates, by S1 nuclease mapping and by hybridization analysis of the in vitro products. The ability of the crude extracts to promote faithful transcription of mouse rDNA correlates with the proliferation rate of the cells. Only extracts prepared from exponentially growing mouse cells contain the factor(s) required for the faithful transcription of mouse ribosomal genes. Extracts from nongrowing or slowly growing mouse cells show very little activity. The cell-free system somehow reflects the rRNA synthetic activity of the cell and will prove valuable for the identification and purification of the various factors that are involved in the specific read-out of rDNA, and may play a central role in the regulation of transcription of the ribosomal genes.This publication has 32 references indexed in Scilit:
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