cis- and trans-Acting determinants of transcription termination by yeast RNA polymerase II.
Most eukaryotic genes are transcribed by RNA polymerase II (Pol II), including those that produce mRNAs and many noncoding functional RNAs. Proper expression of these genes requires efficient termination by Pol II to avoid transcriptional interference and synthesis of extended, nonfunctional RNAs. We previously described a pathway for yeast Pol II termination that involves recognition of an element in the nascent transcript by the essential RNA-binding protein Nrd1. The Nrd1-dependent pathway appears to be used primarily for nonpolyadenylated transcripts, such as the small nuclear and small nucleolar RNAs (snoRNAs). mRNAs are thought to use a distinct pathway that is coupled to cleavage and polyadenylation of the transcript. Here we show that the terminator elements for two yeast snoRNA genes also direct polyadenylated 3'-end formation in the context of an mRNA 3' untranslated region. A selection for cis-acting terminator readthrough mutations identified conserved features of these elements, some of which are similar to cleavage and polyadenylation signals. A selection for trans-acting mutations that induce readthrough of both a snoRNA and an mRNA terminator yielded mutations in the Rpb3 and Rpb11 subunits of Pol II that define a remarkably discrete surface on the trailing end of the enzyme. Our results suggest that, at least in budding yeast, protein-coding and noncoding Pol II-transcribed genes use similar mechanisms to direct termination and that the termination signal is transduced through the Rpb3/Rpb11 heterodimer.
Department of Biomolecular Chemistry, University of Wisconsin Medical School, 1300 University Avenue, Madison, Wisconsin 53706-1532, USA.
Eric J
EJ
Steinmetz
Sarah B H
SB
Ng
Joseph P
JP
Cloute
David A
DA
Brow
D000595:Amino Acid Sequence; D001483:Base Sequence; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D026723:Polyadenylation / Q000235:genetics; D021122:Protein Subunits / Q000378:metabolism; D012319:RNA Polymerase II / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D020537:RNA, Small Nucleolar / Q000235:genetics; D050436:Regulatory Elements, Transcriptional / Q000235:genetics*; D013728:Terminator Regions, Genetic / Q000235:genetics*; D015534:Trans-Activators / Q000235:genetics*; D014158:Transcription, Genetic / Q000235:genetics*; D015003:Yeasts / Q000201:enzymology* / Q000235:genetics*
D021122:Protein Subunits; D012333:RNA, Messenger; D020537:RNA, Small Nucleolar; D015534:Trans-Activators; D012319:RNA Polymerase II