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Nucleic Acids Research, 1990, Vol. 18, No. 7 1677-1718
© 1990

MOLECULAR BIOLOGY

Interaction of RNA polymerase I transcription factors with a promoter in the nontranscribed spacer of rat ribosomal DNA

S.David Smith, Emmanuel Oriahi, Hsin-Fang Yang-Yen, WenQin Xie, Catherine Chen and Lawrence I. Rothblum*

Weis Center for Research, Geisinger Clinic Danville, PA 17822, USA

* To whom correspondence should be addressed

Received January 29, 1990. Revised March 1, 1990. Accepted March 1, 1990.

The spacer promoter of the rat rDNA repeat consists of two functional domains: a core (proximal) element that is sufficient for transcription In vitro, and an upstream (distal) promoter element that increases the efficiency of transcription. Two of the transcription factors that interact with the 45S promoter also interact with the spacer promoter. Rat SL-1, is required for transcription of the spacer promoter by heterologous extracts, e.g. human, and rat SF-1 is required for efficient transcription in vitro. Order-of-addition experiments demonstrated that the preinitiation complex formed by these factors on the spacer promotor Is not as stable as the complex formed on the 45S promoter. DNase 1 footprlnting experiments demonstrated binding sites for rat SL-1 and SF-1 on the distal element of the spacer promoter. The topology of the domains of the spacer promoter may explain both the reduced stability of the preinitiation complex . formed on that promoter and the lower efficiency of transcription of that promo ter when compared to the 45S promoter


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