Free poly(A) stimulates capped mRNA translation in vitro through the eIF4G-poly(A)-binding protein interaction

J Biol Chem. 2002 Sep 27;277(39):36818-24. doi: 10.1074/jbc.M205065200. Epub 2002 Jul 22.

Abstract

The 5' cap and 3' poly(A) tail of classical eukaryotic mRNAs functionally communicate to synergistically enhance translation initiation. Synergy has been proposed to result in part from facilitated ribosome recapture on circularized mRNAs. Here, we demonstrate that this is not the case. In poly(A)-dependent, ribosome-depleted rabbit reticulocyte lysates, the addition of exogenous poly(A) chains of physiological length dramatically stimulated translation of a capped, nonpolyadenylated mRNA. When the poly(A):RNA ratio approached 1, exogenous poly(A) stimulated translation to the same extent as the presence of a poly(A) tail at the mRNA 3' end. In addition, exogenous poly(A) significantly improved translation of capped mRNAs carrying short poly(A(50)) tails. Trans stimulation of translation by poly(A) required the eIF4G-poly(A)-binding protein interaction and resulted in increased affinity of eIF4E for the mRNA cap, exactly as we recently described for cap-poly(A) synergy. These results formally demonstrate that mRNA circularization per se is not the cause of cap-poly(A) synergy at least in vitro.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Binding, Competitive
  • Dose-Response Relationship, Drug
  • Escherichia coli / metabolism
  • Eukaryotic Initiation Factor-4G / chemistry
  • Eukaryotic Initiation Factor-4G / metabolism*
  • Models, Biological
  • Plasmids / metabolism
  • Poly A / metabolism*
  • Poly(A)-Binding Proteins / chemistry
  • Poly(A)-Binding Proteins / metabolism*
  • Polyadenylation
  • Protein Binding
  • Protein Biosynthesis*
  • RNA, Messenger / metabolism*
  • RNA, Messenger / pharmacology
  • Ribosomes / metabolism
  • Transcription, Genetic
  • Xenopus

Substances

  • Eukaryotic Initiation Factor-4G
  • Poly(A)-Binding Proteins
  • RNA, Messenger
  • Poly A