Saccharomyces cerevisiae RNA polymerase I terminates transcription at the Reb1 terminator in vivo.
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.
Reeder
R H
RH
Fred Hutchinson Cancer Research Center, Seattle, Washington 98109, USA. rreeder@fred.fhcrc.org
Guevara
P
P
Roan
J G
JG
D001483:Base Sequence; D015342:DNA Probes; D004268:DNA-Binding Proteins / Q000235:genetics*; D004722:Endoribonucleases / Q000378:metabolism; D005800:Genes, Fungal; D008969:Molecular Sequence Data; D009154:Mutation; D011485:Protein Binding; D012318:RNA Polymerase I / Q000378:metabolism*; D012323:RNA Processing, Post-Transcriptional; D012335:RNA, Ribosomal / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D043244:Ribonuclease III; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins; D013728:Terminator Regions, Genetic; D014157:Transcription Factors; D014158:Transcription, Genetic
D015342:DNA Probes; D004268:DNA-Binding Proteins; C065681:REB1 protein, S cerevisiae; D012335:RNA, Ribosomal; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; C065039:RNA, ribosomal, 25S; D012318:RNA Polymerase I; D004722:Endoribonucleases; C468367:RNT1 protein, S cerevisiae; D043244:Ribonuclease III