Genetic and pharmacological evidence for kinetic competition between alternative poly(A) sites in yeast

Elife. 2021 Jul 7:10:e65331. doi: 10.7554/eLife.65331.

Abstract

Most eukaryotic mRNAs accommodate alternative sites of poly(A) addition in the 3' untranslated region in order to regulate mRNA function. Here, we present a systematic analysis of 3' end formation factors, which revealed 3'UTR lengthening in response to a loss of the core machinery, whereas a loss of the Sen1 helicase resulted in shorter 3'UTRs. We show that the anti-cancer drug cordycepin, 3' deoxyadenosine, caused nucleotide accumulation and the usage of distal poly(A) sites. Mycophenolic acid, a drug which reduces GTP levels and impairs RNA polymerase II (RNAP II) transcription elongation, promoted the usage of proximal sites and reversed the effects of cordycepin on alternative polyadenylation. Moreover, cordycepin-mediated usage of distal sites was associated with a permissive chromatin template and was suppressed in the presence of an rpb1 mutation, which slows RNAP II elongation rate. We propose that alternative polyadenylation is governed by temporal coordination of RNAP II transcription and 3' end processing and controlled by the availability of 3' end factors, nucleotide levels and chromatin landscape.

Keywords: S. cerevisiae; Sen1; alternative polyadenylation; chromosomes; cleavage; cordycepin; gene expression; genetics; genomics; mycophenolic acid; polyadenylation; transcription elongation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 3' Untranslated Regions
  • DNA Helicases
  • Kinetics
  • Poly A / chemistry*
  • Polyadenylation*
  • RNA Helicases
  • Saccharomyces cerevisiae / chemistry
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae / metabolism*
  • Saccharomyces cerevisiae Proteins

Substances

  • 3' Untranslated Regions
  • Saccharomyces cerevisiae Proteins
  • Poly A
  • SEN1 protein, S cerevisiae
  • DNA Helicases
  • RNA Helicases

Associated data

  • GEO/GSE160539