The mechanism of action of initiaton factor 3 in protein synthesis,I. Studies on ribosomes crosslinked with dimethylsuberimidate |
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Authors: | David A Hawley Martha J Miller Lawrence I Slobin Albert J Wahba |
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Institution: | 1. Laboratory of Molecular Biology, University of Sherbrooke School of Medicine, Sherbrooke, Quebec, Canada;2. Laboratory for Physiological Chemistry, State University of Leiden, Leiden, The Netherlands |
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Abstract: | The mechanism of action of chain initiation factor 3 in translation was examined by using 70S ribosomes which were covalently crosslinked with dimethylsuberimidate. Crosslinked ribosomes were inactive in AUG-dependent fMet-tRNA binding, and were not stimulated by IF-3 in poly(U) translation. IF-3 is known to be required for maximal rates of amino acid incorporation with synthetic polynucleotides at 18 mM Mg2+. A direct interaction of IF-3 with 70S ribosomes was demonstrated by crosslinking 14C-labeled IF-3 to 70S ribosomes. The labeled factor was also crosslinked to 30S and 50S ribosomal subunits. A model is presented proposing the mechanism of action of IF-3 on 70S ribosomes. |
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