title: Pcf11 orchestrates transcription termination pathways in yeast.
abstract: In Saccharomyces cerevisiae, short noncoding RNA (ncRNA) generated by RNA polymerase II (Pol II) are terminated by the NRD complex consisting of Nrd1, Nab3, and Sen1. We now show that Pcf11, a component of the cleavage and polyadenylation complex (CPAC), is also generally required for NRD-dependent transcription termination through the action of its C-terminal domain (CTD)-interacting domain (CID). Pcf11 localizes downstream from Nrd1 on NRD terminators, and its recruitment depends on Nrd1. Furthermore, mutation of the Pcf11 CID results in Nrd1 retention on chromatin, delayed degradation of ncRNA, and restricted Pol II CTD Ser2 phosphorylation and Sen1-Pol II interaction. Finally, the pcf11-13 and sen1-1 mutant phenotypes are very similar, as both accumulate RNA:DNA hybrids and display Pol II pausing downstream from NRD terminators. We predict a mechanism by which the exchange of Nrd1 and Pcf11 on chromatin facilitates Pol II pausing and CTD Ser2-P phosphorylation. This in turn promotes Sen1 activity that is required for NRD-dependent transcription termination in vivo.
authors: Sir William Dunn School of Pathology, University of Oxford, Oxford OX1 3RE, United Kingdom;
authors: Pawel
authors: P
authors: Grzechnik
authors: Department of Biochemistry, University of Oxford, Oxford OX1 3QU, United Kingdom.
authors: Michal Ryszard
authors: MR
authors: Gdula
authors: Sir William Dunn School of Pathology, University of Oxford, Oxford OX1 3RE, United Kingdom; nicholas.proudfoot@path.ox.ac.uk.
authors: Nick J
authors: NJ
authors: Proudfoot
mesh_terms: D004265:DNA Helicases / Q000235:genetics; D017434:Protein Structure, Tertiary; D020365:RNA Helicases / Q000235:genetics; D022661:RNA, Untranslated / Q000235:genetics; D016601:RNA-Binding Proteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D061806:Transcription Termination, Genetic / Q000502:physiology*; D039221:mRNA Cleavage and Polyadenylation Factors / Q000235:genetics / Q000378:metabolism*
chemical_list: C491211:NRD1 protein, S cerevisiae; C512637:PCF11 protein, S cerevisiae; D022661:RNA, Untranslated; D016601:RNA-Binding Proteins; D029701:Saccharomyces cerevisiae Proteins; D039221:mRNA Cleavage and Polyadenylation Factors; C542347:SEN1 protein, S cerevisiae; D004265:DNA Helicases; D020365:RNA Helicases
keywords: CTD phosphorylation; NRD complex; cleavage and polyadenylation complex; noncoding RNA; transcription termination