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Eukaryotic ribosomes require initiation factors 1 and 1A to locate initiation codons

Nature volume 394, pages 854–859 (1998)Cite this article

Abstract

The scanning model of translation initiation is a coherent description of how eukaryotic ribosomes reach the initiation codon after being recruited to the capped 5′ end of messenger RNA. Five eukaryotic initiation factors (eIF 2, 3, 4A, 4B and 4F) with established functions have been assumed to be sufficient to mediate this process. Here we report that eIF1 and eIF1A are also both essential for translation initiation. In their absence, 43S ribosomal preinitiation complexes incubated with ATP, eIF4A, eIF4B and eIF4F bind exclusively to the cap-proximal region but are unable to reach the initiation codon. Individually, eIF1A enhances formation of this cap-proximal complex, and eIF1 weakly promotes formation of a 48S ribosomal complex at the initiation codon. These proteins act synergistically to mediate assembly of ribosomal initiation complexes at the initiation codon and dissociate aberrant complexes from the mRNA.

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Figure 1: Composition and purification of proteins used in translation initiation.
Figure 2: Assembly and toeprint analysis of ribosomal complexes on β-globin mRNA.
Figure 3: Initiation-factor dependence of cap-mediated 48S complex formation.
Figure 4: Stability of complex I.
Figure 5: Assembly and toeprint analysis of ribosomal complexes on EMCV and CSFV mRNAs.

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Acknowledgements

We thank W. Merrick and N. Sonenberg for discussion, L. Siconolfi-Baez for sequencing the eIF1 and eIF1A proteins, and H.-P. Vornlocher and J. W. B. Hershey for the pT7-7-1A plasmid. These studies were supported by grants from the NIH (C.U.T.H. and S.I.B.) and the NSF (C.U.T.H.).

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Authors and Affiliations

  1. Department of Microbiology and Immunology, State University of New York Health Science Center at Brooklyn, 450 Clarkson Avenue, Brooklyn, 11203, New York, USA

    Tatyana V. Pestova, Sergei I. Borukhov & Christopher U. T. Hellen

  2. A.N. Belozersky Institute of Physico-Chemical Biology, Moscow State University, 119899, Moscow, Russia

    Tatyana V. Pestova

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  1. Tatyana V. Pestova
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Correspondence to Tatyana V. Pestova.

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Pestova, T., Borukhov, S. & Hellen, C. Eukaryotic ribosomes require initiation factors 1 and 1A to locate initiation codons. Nature 394, 854–859 (1998). https://doi.org/10.1038/29703

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