Uridylation and PABP Cooperate to Repair mRNA Deadenylated Ends in Arabidopsis

Cell Rep. 2016 Mar 22;14(11):2707-17. doi: 10.1016/j.celrep.2016.02.060. Epub 2016 Mar 10.

Abstract

Uridylation emerges as a key modification promoting mRNA degradation in eukaryotes. In addition, uridylation by URT1 prevents the accumulation of excessively deadenylated mRNAs in Arabidopsis. Here, we show that the extent of mRNA deadenylation is controlled by URT1. By using TAIL-seq analysis, we demonstrate the prevalence of mRNA uridylation and the existence, at lower frequencies, of mRNA cytidylation and guanylation in Arabidopsis. Both URT1-dependent and URT1-independent types of uridylation co-exist but only URT1-mediated uridylation prevents the accumulation of excessively deadenylated mRNAs. Importantly, uridylation repairs deadenylated extremities to restore the size distribution observed for non-uridylated oligo(A) tails. In vivo and in vitro data indicate that Poly(A) Binding Protein (PABP) binds to uridylated oligo(A) tails and determines the length of U-extensions added by URT1. Taken together, our results uncover a role for uridylation and PABP in repairing mRNA deadenylated ends and reveal that uridylation plays diverse roles in eukaryotic mRNA metabolism.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arabidopsis / genetics
  • Arabidopsis / metabolism*
  • Arabidopsis Proteins / antagonists & inhibitors
  • Arabidopsis Proteins / genetics
  • Arabidopsis Proteins / metabolism*
  • Binding Sites
  • Blotting, Western
  • Immunoprecipitation
  • MicroRNAs / metabolism
  • Poly A / metabolism
  • Poly U / metabolism
  • Poly(A)-Binding Proteins / genetics
  • Poly(A)-Binding Proteins / metabolism*
  • RNA Interference
  • RNA Nucleotidyltransferases / genetics
  • RNA Nucleotidyltransferases / metabolism*
  • RNA Stability
  • RNA, Messenger / metabolism*
  • RNA, Small Interfering / metabolism
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / isolation & purification

Substances

  • Arabidopsis Proteins
  • MicroRNAs
  • Poly(A)-Binding Proteins
  • RNA, Messenger
  • RNA, Small Interfering
  • Recombinant Proteins
  • Poly A
  • Poly U
  • RNA Nucleotidyltransferases
  • URT1 protein, Arabidopsis