A novel function for eukaryotic elongation factor 3: Inhibition of stop codon readthrough in yeast. 2023

Soushi Kobayashi, and Akira Kaji, and Hideko Kaji
Department of Microbiology, Perelman School of Medicine, University of Pennsylvania, 3610 Hamilton Walk, Philadelphia, PA, 19104, USA; Department of Biochemistry and Molecular Biology, Thomas Jefferson University, 1020 Locust Street, Philadelphia, PA, 19107, USA. Electronic address: soushik@pharm.showa-u.ac.jp.

Eukaryotic elongation factor 3 (eEF3) is one of the essential yeast ribosome-associated ATP-binding cassette type F (ABCF) ATPases. Previously, we found that eEF3 stimulates release of mRNA from puromycin-treated polysomes. In this study, we used a cell-free cricket paralysis virus (CrPV) internal ribosome entry site (IRES)-mediated firefly luciferase bicistronic mRNA translation system with yeast S30 extract. When eEF3 was partially removed from the crude extract, the product from the downstream ORF was increased by the readthrough of a UAA stop codon in the upstream ORF. eEF3 enhanced the release of luciferase from the polysome by eukaryotic release factor (eRF)1 and eRF3. These results suggest that eEF3 is a factor that assists eRFs in performing normal protein synthesis termination in yeast.

UI MeSH Term Description Entries
D010445 Peptide Elongation Factors Protein factors uniquely required during the elongation phase of protein synthesis. Elongation Factor,Elongation Factors, Peptide,Factor, Elongation,Factors, Peptide Elongation
D012270 Ribosomes Multicomponent ribonucleoprotein structures found in the CYTOPLASM of all cells, and in MITOCHONDRIA, and PLASTIDS. They function in PROTEIN BIOSYNTHESIS via GENETIC TRANSLATION. Ribosome
D012441 Saccharomyces cerevisiae A species of the genus SACCHAROMYCES, family Saccharomycetaceae, order Saccharomycetales, known as "baker's" or "brewer's" yeast. The dried form is used as a dietary supplement. Baker's Yeast,Brewer's Yeast,Candida robusta,S. cerevisiae,Saccharomyces capensis,Saccharomyces italicus,Saccharomyces oviformis,Saccharomyces uvarum var. melibiosus,Yeast, Baker's,Yeast, Brewer's,Baker Yeast,S cerevisiae,Baker's Yeasts,Yeast, Baker
D014176 Protein Biosynthesis The biosynthesis of PEPTIDES and PROTEINS on RIBOSOMES, directed by MESSENGER RNA, via TRANSFER RNA that is charged with standard proteinogenic AMINO ACIDS. Genetic Translation,Peptide Biosynthesis, Ribosomal,Protein Translation,Translation, Genetic,Protein Biosynthesis, Ribosomal,Protein Synthesis, Ribosomal,Ribosomal Peptide Biosynthesis,mRNA Translation,Biosynthesis, Protein,Biosynthesis, Ribosomal Peptide,Biosynthesis, Ribosomal Protein,Genetic Translations,Ribosomal Protein Biosynthesis,Ribosomal Protein Synthesis,Synthesis, Ribosomal Protein,Translation, Protein,Translation, mRNA,mRNA Translations
D018388 Codon, Terminator Any codon that signals the termination of genetic translation (TRANSLATION, GENETIC). PEPTIDE TERMINATION FACTORS bind to the stop codon and trigger the hydrolysis of the aminoacyl bond connecting the completed polypeptide to the tRNA. Terminator codons do not specify amino acids. Amber Stop Codon,Codon, Amber Stop,Codon, Ochre Stop,Codon, Opal Stop,Codon, Stop,Stop Codon,Terminator Codon,Amber Codon,Amber Terminator Codon,Codon, Termination,Ochre Codon,Ochre Stop Codon,Opal Codon,Opal Stop Codon,Stop Codon UAA,Stop Codon UAG,Stop Codon UGA,Stop Signal, Translation,TAA Codon,TAG Codon,TGA Codon,UAA Codon,UAA Stop Codon,UAG Codon,UAG Stop Codon,UGA Codon,UGA Stop Codon,Amber Codons,Amber Stop Codons,Amber Terminator Codons,Codon UAA, Stop,Codon UAG, Stop,Codon UGA, Stop,Codon, Amber,Codon, Amber Terminator,Codon, Ochre,Codon, Opal,Codon, TAA,Codon, TAG,Codon, TGA,Codon, UAA,Codon, UAA Stop,Codon, UAG,Codon, UAG Stop,Codon, UGA,Codon, UGA Stop,Codons, Amber,Codons, Amber Stop,Codons, Amber Terminator,Codons, Ochre,Codons, Ochre Stop,Codons, Opal,Codons, Opal Stop,Codons, Stop,Codons, TAA,Codons, TAG,Codons, TGA,Codons, Termination,Codons, Terminator,Codons, UAA,Codons, UAA Stop,Codons, UAG,Codons, UAG Stop,Codons, UGA,Codons, UGA Stop,Ochre Codons,Ochre Stop Codons,Opal Codons,Opal Stop Codons,Stop Codon UGAs,Stop Codon, Amber,Stop Codon, Ochre,Stop Codon, Opal,Stop Codon, UAA,Stop Codon, UAG,Stop Codon, UGA,Stop Codons,Stop Codons, Amber,Stop Codons, Ochre,Stop Codons, Opal,Stop Codons, UAA,Stop Codons, UAG,Stop Codons, UGA,Stop Signals, Translation,TAA Codons,TAG Codons,TGA Codons,Termination Codon,Termination Codons,Terminator Codon, Amber,Terminator Codons,Terminator Codons, Amber,Translation Stop Signal,Translation Stop Signals,UAA Codons,UAA Stop Codons,UAA, Stop Codon,UAG Codons,UAG Stop Codons,UAG, Stop Codon,UGA Codons,UGA Stop Codons,UGA, Stop Codon

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