Possible steps of complete disassembly of post-termination complex by yeast eEF3 deduced from inhibition by translocation inhibitors

Shinya Kurata, Ben Shen, Jun Liu, Nono Takeuchi, Akira Kaji, Hideko Kaji

Research output: Contribution to journalArticle

Abstract

Ribosomes, after one round of translation, must be recycled so that the next round of translation can occur. Complete disassembly of post-termination ribosomal complex (PoTC) in yeast for the recycling consists of three reactions: release of tRNA, release of mRNA and splitting of ribosomes, catalyzed by eukaryotic elongation factor 3 (eEF3) and ATP. Here, we show that translocation inhibitors cycloheximide and lactimidomycin inhibited all three reactions. Cycloheximide is a non-competitive inhibitor of both eEF3 and ATP. The inhibition was observed regardless of the way PoTC was prepared with either release factors or puromycin. Paromomycin not only inhibited all three reactions but also re-associated yeast ribosomal subunits. On the other hand, sordarin or fusidic acid, when applied together with eEF2/GTP, specifically inhibited ribosome splitting without blocking of tRNA/mRNA release. From these inhibitor studies, we propose that, in accordance with eEF3's known function in elongation, the release of tRNA via exit site occurs first, then mRNA is released, followed by the splitting of ribosomes during the disassembly of post-termination complexes catalyzed by eEF3 and ATP.

Original languageEnglish (US)
Pages (from-to)264-276
Number of pages13
JournalNucleic Acids Research
Volume41
Issue number1
DOIs
StatePublished - Jan 2013

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Peptide Elongation Factors
Ribosomes
Transfer RNA
Yeasts
Adenosine Triphosphate
Cycloheximide
Messenger RNA
Paromomycin
Fusidic Acid
Ribosome Subunits
Puromycin
Recycling
Guanosine Triphosphate

ASJC Scopus subject areas

  • Genetics

Cite this

Possible steps of complete disassembly of post-termination complex by yeast eEF3 deduced from inhibition by translocation inhibitors. / Kurata, Shinya; Shen, Ben; Liu, Jun; Takeuchi, Nono; Kaji, Akira; Kaji, Hideko.

In: Nucleic Acids Research, Vol. 41, No. 1, 01.2013, p. 264-276.

Research output: Contribution to journalArticle

Kurata, Shinya ; Shen, Ben ; Liu, Jun ; Takeuchi, Nono ; Kaji, Akira ; Kaji, Hideko. / Possible steps of complete disassembly of post-termination complex by yeast eEF3 deduced from inhibition by translocation inhibitors. In: Nucleic Acids Research. 2013 ; Vol. 41, No. 1. pp. 264-276.
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