Abstract
The translation initiation factor eIF1A is necessary for directing the 43S preinitiation complex from the 5' end of the mRNA to the initiation codon in a process termed scanning. We have determined the solution structure of human eIF1A, which reveals an oligonucleotide-binding (OB) fold and an additional domain. NMR titration experiments showed that eIF1A binds single-stranded RNA oligonucleotides in a site-specific, but non-sequence-specific manner, hinting at an mRNA interaction rather than specific rRNA or tRNA binding. The RNA binding surface extends over a large area covering the canonical OB fold binding site as well as a groove leading to the second domain. Site-directed mutations at multiple positions along the RNA-binding surface were defective in the ability to properly assemble preinitiation complexes at the AUG codon in vitro.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Amino Acid Sequence
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Archaea
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Bacteria
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Base Sequence
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Binding Sites
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Codon
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Eukaryotic Initiation Factor-1*
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Humans
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Models, Molecular
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Molecular Sequence Data
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Nuclear Magnetic Resonance, Biomolecular
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Oligoribonucleotides / chemistry*
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Oligoribonucleotides / metabolism
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Peptide Initiation Factors / chemistry*
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Peptide Initiation Factors / metabolism*
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Protein Folding
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Protein Structure, Secondary
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RNA, Messenger / chemistry
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RNA, Messenger / metabolism*
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Recombinant Proteins / chemistry
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Recombinant Proteins / metabolism
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Saccharomyces cerevisiae
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Sequence Alignment
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Sequence Homology, Amino Acid
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Solutions
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Substrate Specificity
Substances
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Codon
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Eukaryotic Initiation Factor-1
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Oligoribonucleotides
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Peptide Initiation Factors
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RNA, Messenger
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Recombinant Proteins
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Solutions
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eukaryotic peptide initiation factor-1A