Distinct functions of eukaryotic translation initiation factors eIF1A and eIF3 in the formation of the 40 S ribosomal preinitiation complex

Jayanta Chaudhuri, Dipanjan Chowdhury, Umadas Maitra

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Abstract

We have used an in vitro translation initiation assay to investigate the requirements for the efficient transfer of Met-tRNA(f) (as Met- tRNA(f)·eIF2·GTP ternary complex) to 40 S ribosomal subunits in the absence of mRNA (or an AUG codon) to form the 40 S preinitiation complex. We observed that the 17-kDa initiation factor eIF1A is necessary and sufficient to mediate nearly quantitative transfer of Met-tRNA(f) to isolated 40 S ribosomal subunits. However, the addition of 60 S ribosomal subunits to the 40 S preinitiation complex formed under these conditions disrupted the 40 S complex resulting in dissociation of Met-tRNA(f) from the 40 S subunit. When the eIF1A-dependent preinitiation reaction was carried out with 40 S ribosomal subunits that had been preincubated with eIF3, the 40 S preinitiation complex formed included bound eIF3 (40 S·IF3·Met- tRNA(f)·eIF2·GTP). In contrast to the complex lacking eIF3, this complex was not disrupted by the addition of 60 S ribosomal subunits. These results suggest that in vivo, both eIF1A and eIF3 are required to form a stable 40 S preinitiation complex, eIF1A catalyzing the transfer of Met- tRNA(f)·eIF2·GTP to 40 S subunits, and eIF3 stabilizing the resulting complex and preventing its disruption by 60 S ribosomal subunits.

Original languageEnglish (US)
Pages (from-to)17975-17980
Number of pages6
JournalJournal of Biological Chemistry
Volume274
Issue number25
DOIs
StatePublished - Jun 18 1999

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Eukaryotic Initiation Factors
Ribosome Subunits
Peptide Initiation Factors
Transfer RNA
Assays
Codon
Messenger RNA
formylmethionine-tRNA

ASJC Scopus subject areas

  • Biochemistry

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Distinct functions of eukaryotic translation initiation factors eIF1A and eIF3 in the formation of the 40 S ribosomal preinitiation complex. / Chaudhuri, Jayanta; Chowdhury, Dipanjan; Maitra, Umadas.

In: Journal of Biological Chemistry, Vol. 274, No. 25, 18.06.1999, p. 17975-17980.

Research output: Contribution to journalArticle

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