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Molecular and Cellular Biology, November 1999, p. 7369-7376, Vol. 19, No. 11
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Saccharomyces cerevisiae RNA Polymerase I Terminates Transcription at the Reb1 Terminator In Vivo

Ronald H. Reeder,1,* Palmira Guevara,2 and Judith G. Roan1

Fred Hutchinson Cancer Research Center, Seattle, Washington 98109,1 and Grupo de Genética Molecular, Universidad Central de Venezuela, Caracas 1041A, Venezuela2

Received 11 January 1999/Returned for modification 18 February 1999/Accepted 30 July 1999

We have mapped transcription termination sites for RNA polymerase I in the yeast Saccharomyces cerevisiae. S1 nuclease mapping shows that the primary terminator is the Reb1p terminator located at +93 downstream of the 3' end of 25S rRNA. Reverse transcription coupled with quantitative PCR shows that approximately 90% of all transcripts terminate at this site. Transcripts which read through the +93 site quantitatively terminate at a fail-safe terminator located further downstream at +250. Inactivation of Rnt1p (an RNase III involved in processing the 3' end of 25S rRNA) greatly stabilizes transcripts extending to both sites and increases readthrough at the +93 site. In vivo assay of mutants of the Reb1p terminator shows that this site operates in vivo by the same mechanism as has previously been delineated through in vitro studies.

* Corresponding author. Mailing address: Fred Hutchinson Cancer Research Center, 1100 Fairview Ave. N., Seattle, WA 98109. Phone: (206) 667-4513. Fax: (206) 667-4082. E-mail: rreeder{at}fred.fhcrc.org.

Molecular and Cellular Biology, November 1999, p. 7369-7376, Vol. 19, No. 11
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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