A novel role of the mammalian GSPT/eRF3 associating with poly(A)-binding protein in Cap/Poly(A)-dependent translation

J Biol Chem. 2002 Dec 27;277(52):50286-92. doi: 10.1074/jbc.M203029200. Epub 2002 Oct 14.

Abstract

The mammalian GSPT, which consists of amino-terminal (N) and carboxyl-terminal (C) domains, functions as the eukaryotic releasing factor 3 (eRF3) by interacting with eRF1 in translation termination. This function requires only the C-domain that is homologous to the elongation factor (EF) 1alpha, while the N-domain interacts with polyadenylate-binding protein (PABP), which binds the poly(A) tail of mRNA and associates with the eukaryotic initiation factor (eIF) 4G. Here we describe a novel role of GSPT in translation. We first determined an amino acid sequence required for the PABP interaction in the N-domain. Inhibition of this interaction significantly attenuated translation of capped/poly(A)-tailed mRNA not only in an in vitro translation system but also in living cells. There was a PABP-dependent linkage between the termination factor complex eRF1-GSPT and the initiation factor eIF4G associating with 5' cap through eIF4E. Although the inhibition of the GSPT-PABP interaction did not affect the de novo formation of an 80 S ribosomal initiation complex, it appears to suppress the subsequent recycle of ribosome. These results indicate that GSPT/eRF3 plays an important role in translation cycle through the interaction with PABP, in addition to mediating the termination with eRF1.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Binding Sites
  • COS Cells
  • Chlorocebus aethiops
  • Glutathione Transferase / genetics
  • Glutathione Transferase / metabolism
  • HeLa Cells
  • Humans
  • Kinetics
  • Mutagenesis
  • Peptide Termination Factors / chemistry
  • Peptide Termination Factors / genetics
  • Peptide Termination Factors / metabolism*
  • Poly(A)-Binding Proteins / chemistry
  • Poly(A)-Binding Proteins / metabolism*
  • Protein Biosynthesis*
  • Recombinant Fusion Proteins / metabolism
  • Sequence Deletion
  • Transfection

Substances

  • Peptide Termination Factors
  • Poly(A)-Binding Proteins
  • Recombinant Fusion Proteins
  • peptide-chain-release factor 3
  • Glutathione Transferase