Forgot Password

If you have forgotten your password you can enter your email here and get a temporary password sent to your email.

The indispensable N-terminal half of eIF3j/HCR1 cooperates with its structurally conserved binding partner eIF3b/PRT1-RRM and with eIF1A in stringent AUG selection.

Despite recent progress in our understanding of the numerous functions of individual subunits of eukaryotic translation initiation factor (eIF) 3, little is known on the molecular level. Using NMR spectroscopy, we determined the first solution structure of an interaction between eIF3 subunits. We revealed that a conserved tryptophan residue in the human eIF3j N-terminal acidic motif (NTA) is held in the helix alpha1 and loop 5 hydrophobic pocket of the human eIF3b RNA recognition motif (RRM). Mutating the corresponding "pocket" residues in its yeast orthologue reduces cellular growth rate, eliminates eIF3j/HCR1 association with eIF3b/PRT1 in vitro and in vivo, affects 40S occupancy of eIF3, and produces a leaky scanning defect indicative of a deregulation of the AUG selection process. Unexpectedly, we found that the N-terminal half of eIF3j/HCR1 containing the NTA is indispensable and sufficient for wild-type growth of yeast cells. Furthermore, we demonstrate that deletion of either j/HCR1 or its N-terminal half only, or mutation of the key tryptophan residues results in the severe leaky scanning phenotype partially suppressible by overexpressed eIF1A, which is thought to stabilize properly formed preinitiation complexes at the correct start codon. These findings indicate that eIF3j/HCR1 remains associated with the scanning preinitiation complexes and does not dissociate from the small ribosomal subunit upon mRNA recruitment, as previously believed. Finally, we provide further support for earlier mapping of the ribosomal binding site for human eIF3j by identifying specific interactions of eIF3j/HCR1 with small ribosomal proteins RPS2 and RPS23 located in the vicinity of the mRNA entry channel. Taken together, we propose that eIF3j/HCR1 closely cooperates with the eIF3b/PRT1 RRM and eIF1A on the ribosome to ensure proper formation of the scanning-arrested conformation required for stringent AUG recognition.

Pubmed ID: 20060839


  • Elantak L
  • Wagner S
  • Herrmannová A
  • Karásková M
  • Rutkai E
  • Lukavsky PJ
  • Valásek L


Journal of molecular biology

Publication Data

March 5, 2010

Associated Grants

  • Agency: Wellcome Trust, Id: 076456
  • Agency: Wellcome Trust, Id: 076456/Z/05/Z
  • Agency: Medical Research Council, Id: MC_U105170649
  • Agency: FIC NIH HHS, Id: R01 TW007271
  • Agency: FIC NIH HHS, Id: R01 TW007271-01
  • Agency: Medical Research Council, Id:

Mesh Terms

  • Amino Acid Sequence
  • Binding Sites
  • Codon, Initiator
  • Conserved Sequence
  • Eukaryotic Initiation Factor-3
  • Evolution, Molecular
  • Humans
  • In Vitro Techniques
  • Models, Molecular
  • Molecular Sequence Data
  • Multiprotein Complexes
  • Mutagenesis, Site-Directed
  • Nuclear Magnetic Resonance, Biomolecular
  • Peptide Initiation Factors
  • Protein Biosynthesis
  • Protein Interaction Domains and Motifs
  • Protein Subunits
  • Recombinant Proteins
  • Saccharomyces cerevisiae
  • Saccharomyces cerevisiae Proteins
  • Sequence Homology, Amino Acid
  • Tryptophan