Document Type

Article

Publication Date

6-15-2010

Comments

This article has been peer reviewed. It is the authors' final version prior to publication in Proceedings of the National Academy of Sciences of the United States of America Volume 107, Issue 24, 15 June 2010, Pages 10854-10859 The published version is available at DOI: 10.1073/pnas.1006247107. Copyright © National Academy of Sciences

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 new round 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 5gamma'-(beta,gamma-imido)triphosphate did not function at all. The k(cat) 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 on our 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.

Kurata_etal_SI_Text_final.doc (101 kB)
Supporting information

Kurata_etal_SI_Legends_final.doc (43 kB)
Supporting Figures Legend

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Figure S7

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Figure S8

PubMed ID

20534490

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