Abstract
The Saccharomyces cerevisiae TIF3 gene encodes the yeast homologue of mammalian translation initiation factor eIF-4B. We have added six histidine residues to the C-terminus of Tif3 protein (Tif3-His6p) and purified the tagged protein by affinity chromatography. Tif3-His6p stimulates translation and mRNA binding to ribosomes in a Tif3-dependent in vitro system. Furthermore, it binds to single-stranded RNA and catalyses the annealing of partially complementary RNA strands in vitro. In parallel experiments, RNA annealing activity could also be demonstrated for mammalian eIF-4B. A role for Tif3/eIF-4B and RNA annealing activity in the scanning process is proposed.
MeSH terms
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Animals
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Base Sequence
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Eukaryotic Initiation Factors*
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Fungal Proteins / genetics
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Fungal Proteins / metabolism*
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Genes, Fungal / genetics
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Histidine / genetics
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Models, Genetic
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Molecular Sequence Data
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Nucleic Acid Renaturation
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Peptide Chain Initiation, Translational / genetics*
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Peptide Initiation Factors / genetics
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Peptide Initiation Factors / metabolism*
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RNA / metabolism*
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RNA Nucleotidyltransferases / genetics
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RNA Nucleotidyltransferases / metabolism*
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RNA, Messenger / metabolism
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Recombinant Fusion Proteins / biosynthesis
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Recombinant Fusion Proteins / isolation & purification
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Ribosomes / metabolism
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Saccharomyces cerevisiae / genetics*
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Saccharomyces cerevisiae Proteins*
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Sequence Deletion
Substances
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Eukaryotic Initiation Factors
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Fungal Proteins
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Peptide Initiation Factors
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RNA, Messenger
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Recombinant Fusion Proteins
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Saccharomyces cerevisiae Proteins
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TIF3 protein, S cerevisiae
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eIF-4B
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Histidine
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RNA
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RNA Nucleotidyltransferases