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Nucleic Acids Research, 1995, Vol. 23, No. 12 2252-2258
© 1995

MOLECULAR BIOLOGY

The Xenopus 9 bp ribosomal terminator (T3 box) is a pause signal for the RNA polymerase I elongation complex

Paul Labhart

The Scripps Research Institute, Department of Molecular and Experimental Medicine 10666 North Torrey Pines Road, La Jolla, CA 92037, USA

Received February 2, 1995. Revised April 28, 1995. Accepted April 28, 1995.

In Xenopus, termination by RNA polymerase I (pol I) is mediated by the 9 bp sequence GACTTGCNC and RNA 3'-ends are formed –15–20 nt upstream of this terminator element Here I show that this 9 bp element, also called the ‘T3 box’, Is a pause signal for the elongating transcription complex. The two major transcripts in the paused complex have 3'-ends mapping to 15 and 21 nt upstream of the T3 box, remain bound to the template in 0.25' Sarkosyl, are subject to pyrophosphorolysls and can be chased into longer transcripts. Mutations that reduce overall termination also affect pausing, indicating that pausing is a limiting step in the termination process. Oligonucleotide competition experiments, furthermore, suggest that pausing requires a DNA binding factor. The data support a model in which the first step leading to transcription termination by pol I in Xenopus is pausing of the elongation complex upstream of the T3 box.


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