Ribosome recycling step in yeast cytoplasmic protein synthesis is catalyzed by eEF3 and ATP

Shinya Kurata, Klaus H. Nielsen, Sarah F. Mitchell, Jon R. Lorsch, Akira Kaji, Hideko Kaji

Research output: Contribution to journalArticle

Abstract

After each round of protein biosynthesis, the posttermination complex (PoTC) consisting of a ribosome, mRNA, and tRNA must be disassembled into its components for a newround of translation. Here, we show that a Saccharomyces cerevisiae model PoTC was disassembled by ATP and eukaryotic elongation factor 3 (eEF3). GTP or ITP functioned with less efficiency and adenosine 5γ′-(β,γ-imido)triphosphate did not function at all. The kcat of eEF3 was 1.12 min-1, which is comparable to that of the in vitro initiation step. The disassembly reaction was inhibited by aminoglycosides and cycloheximide. The subunits formed from the yeast model PoTC remained separated under ionic conditions close to those existing in vivo, suggesting that they are ready to enter the initiation process. Based onour experimental techniques used in this paper, the release of mRNA and tRNA and ribosome dissociation took place simultaneously. No 40S•mRNA complex was observed, indicating that eEF3 action promotes ribosome recycling, not reinitiation.

Original languageEnglish (US)
Pages (from-to)10854-10859
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume107
Issue number24
DOIs
StatePublished - Jun 15 2010

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Peptide Elongation Factors
Fungal Proteins
Ribosomes
Adenosine Triphosphate
Transfer RNA
Inosine Triphosphate
Messenger RNA
Protein Biosynthesis
Aminoglycosides
Cycloheximide
Guanosine Triphosphate
Adenosine
Saccharomyces cerevisiae
Yeasts

Keywords

  • Antibiotics
  • Kinetics
  • Ribosomal subunits
  • Ribosome recycling factor
  • Yeast cytoplasm

ASJC Scopus subject areas

  • General

Cite this

Ribosome recycling step in yeast cytoplasmic protein synthesis is catalyzed by eEF3 and ATP. / Kurata, Shinya; Nielsen, Klaus H.; Mitchell, Sarah F.; Lorsch, Jon R.; Kaji, Akira; Kaji, Hideko.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 107, No. 24, 15.06.2010, p. 10854-10859.

Research output: Contribution to journalArticle

Kurata, Shinya ; Nielsen, Klaus H. ; Mitchell, Sarah F. ; Lorsch, Jon R. ; Kaji, Akira ; Kaji, Hideko. / Ribosome recycling step in yeast cytoplasmic protein synthesis is catalyzed by eEF3 and ATP. In: Proceedings of the National Academy of Sciences of the United States of America. 2010 ; Vol. 107, No. 24. pp. 10854-10859.
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