Yhh1p/Cft1p directly links poly(A) site recognition and RNA polymerase II transcription termination

EMBO J. 2002 Aug 1;21(15):4125-35. doi: 10.1093/emboj/cdf390.

Abstract

RNA polymerase II (pol II) transcription termination requires co-transcriptional recognition of a functional polyadenylation signal, but the molecular mechanisms that transduce this signal to pol II remain unclear. We show that Yhh1p/Cft1p, the yeast homologue of the mammalian AAUAAA interacting protein CPSF 160, is an RNA-binding protein and provide evidence that it participates in poly(A) site recognition. Interestingly, RNA binding is mediated by a central domain composed of predicted beta-propeller-forming repeats, which occurs in proteins of diverse cellular functions. We also found that Yhh1p/Cft1p bound specifically to the phosphorylated C-terminal domain (CTD) of pol II in vitro and in a two-hybrid test in vivo. Furthermore, transcriptional run-on analysis demonstrated that yhh1 mutants were defective in transcription termination, suggesting that Yhh1p/Cft1p functions in the coupling of transcription and 3'-end formation. We propose that direct interactions of Yhh1p/Cft1p with both the RNA transcript and the CTD are required to communicate poly(A) site recognition to elongating pol II to initiate transcription termination.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actins*
  • Amino Acid Sequence
  • Binding Sites
  • Escherichia coli
  • Macromolecular Substances
  • Molecular Sequence Data
  • Peptide Chain Elongation, Translational
  • Peptide Chain Termination, Translational / physiology*
  • Poly A / metabolism*
  • Protein Structure, Tertiary
  • RNA Polymerase II / physiology*
  • RNA, Fungal / metabolism*
  • RNA, Messenger / metabolism*
  • RNA-Binding Proteins / chemistry
  • RNA-Binding Proteins / genetics
  • RNA-Binding Proteins / physiology*
  • Recombinant Fusion Proteins / physiology
  • Saccharomyces cerevisiae Proteins / chemistry
  • Saccharomyces cerevisiae Proteins / genetics
  • Saccharomyces cerevisiae Proteins / physiology*
  • Sequence Alignment
  • Sequence Homology, Amino Acid
  • Transcription, Genetic
  • Two-Hybrid System Techniques
  • mRNA Cleavage and Polyadenylation Factors

Substances

  • Act1 protein, S cerevisiae
  • Actins
  • Cft1 protein, S cerevisiae
  • Macromolecular Substances
  • RNA, Fungal
  • RNA, Messenger
  • RNA-Binding Proteins
  • Recombinant Fusion Proteins
  • Saccharomyces cerevisiae Proteins
  • mRNA Cleavage and Polyadenylation Factors
  • Poly A
  • RNA Polymerase II