Conversion of 48S translation preinitiation complexes into 80S initiation complexes as revealed by toeprinting

Sergey E. Dmitriev, Andrey V. Pisarev, Maria P. Rubtsova, Yan E. Dunaevsky, Ivan N. Shatsky

Research output: Contribution to journalArticlepeer-review

55 Citations (Scopus)

Abstract

A method of analysis of translation initiation complexes by toeprinting has recently acquired a wide application to investigate molecular mechanisms of translation initiation in eukaryotes. So far, this very fruitful approach was used when researchers did not aim to discriminate between patterns of toeprints for 48S and 80S translation initiation complexes. Here, using cap-dependent and internal ribosomal entry site (IRES)-dependent mRNAs, we show that the toeprint patterns for 48S and 80S complexes are distinct whether the complexes are assembled in rabbit reticulocyte lysate or from fully purified individual components. This observation allowed us to demonstrate for the first time a delay in the conversion of the 48S complex into the 80S complex for β-globin and encephalomyocarditis virus (EMCV) RNAs, and to assess the potential of some 80S antibiotics to block polypeptide elongation. Besides, additional selection of the authentic initiation codon among three consecutive AUGs that follow the EMCV IRES was revealed at steps subsequent to the location of the initiation codon by the 40S ribosomal subunit.

Original languageEnglish
Pages (from-to)99-104
Number of pages6
JournalFEBS Letters
Volume533
Issue number1-3
DOIs
Publication statusPublished - 2 Jan 2003
Externally publishedYes

Keywords

  • AUG selection
  • Eukaryote
  • Toeprint assay
  • Translation initiation

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