Communication between eukaryotic translation initiation factors 1 and 1A on the yeast small ribosomal subunit

David Maag, Jon R. Lorsch

Research output: Contribution to journalArticlepeer-review

Abstract

We have used expressed protein ligation to site-specifically label eukaryotic translation initiation factors (eIFs) 1 and 1A at their C termini with tetramethyl rhodamine. These fluorescent proteins were used in steady-state anisotropy-based binding experiments to measure the dissociation constants of the factors and the yeast small (40S) ribosomal subunit for the first time. These studies demonstrate that both eIF1 and eIF1A are capable of binding to the 40S subunit in the absence of any other initiation factors or mRNA, arguing against previous suggestions that eIF3 is required for recruitment of eIF1 to the small ribosomal subunit. Strikingly, the data also demonstrate that there is approximately ninefold thermodynamic coupling in the binding of the two factors to the 40S subunit. This indicates that eIF1 and eIF1A communicate with one another when bound to the 40S subunit. Communication between these two factors is likely to be important for coordinating their functions during the initiation process. The data presented here provide a foundation on which to build a quantitative understanding of the network of interactions between these essential factors and the rest of the initiation machinery.

Original languageEnglish (US)
Pages (from-to)917-924
Number of pages8
JournalJournal of molecular biology
Volume330
Issue number5
DOIs
StatePublished - Jul 25 2003
Externally publishedYes

Keywords

  • Expressed protein ligation
  • Fluorescence anisotropy
  • Thermodynamic framework

ASJC Scopus subject areas

  • Biophysics
  • Structural Biology
  • Molecular Biology

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