Yeast snoRNA accumulation relies on a cleavage-dependent/polyadenylation-independent 3'-processing apparatus

EMBO J. 2000 Nov 15;19(22):6218-29. doi: 10.1093/emboj/19.22.6218.

Abstract

In Saccharomyces cerevisiae, snoRNAs are encoded by independent genes and within introns. Despite this heterogenous organization, snoRNA biosynthesis relies on a common theme: entry sites for 5'-3' and 3'-5' exonucleases are created on precursor molecules allowing the release of mature snoRNAs. In independently transcribed snoRNAs, such entry sites are often produced by the Rnt1p endonuclease. In many cases, cleavage sites are absent in the 3' portion of the pre-snoRNAs, suggesting that processing starts from the 3' end of the primary transcript. Here we show that cleavage/polyadenylation sites driving efficient polyadenylation, such as CYC1, prevent production of mature and functional snoRNPs. With these sites, snoRNA accumulation is restored only if polyadenylation activity is inhibited. Analysis of sequences downstream of snoRNA-coding units and the use of strains carrying mutations in RNA polymerase II (polII) cleavage/polyadenylation activities allowed us to establish that formation of snoRNA mature 3' ends requires only the cleavage activity of the polII 3'-processing machinery. These data indicate that, in vivo, uncoupling of cleavage and polyadenylation is necessary for an essential cellular biosynthesis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Binding Sites
  • DNA Polymerase II / genetics
  • DNA Polymerase II / metabolism
  • Mutation
  • Nucleic Acid Conformation
  • Oligodeoxyribonucleotides / genetics
  • Poly A / metabolism
  • RNA Processing, Post-Transcriptional
  • RNA, Fungal / chemistry
  • RNA, Fungal / genetics
  • RNA, Fungal / metabolism*
  • RNA, Small Nucleolar / chemistry
  • RNA, Small Nucleolar / genetics
  • RNA, Small Nucleolar / metabolism*
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae / metabolism*
  • Transcription, Genetic

Substances

  • Oligodeoxyribonucleotides
  • RNA, Fungal
  • RNA, Small Nucleolar
  • Poly A
  • DNA Polymerase II