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

Kurata_etal_Fig1.pdf (312 kB)
Figure 1

Kurata_etal_Fig2.pdf (54 kB)
Figure 2

Kurata_etal_Fig3.pdf (140 kB)
Figure 3

Kurata_etal_Fig4.pdf (38 kB)
Figure 4

Kurata_etal_Fig5.pdf (199 kB)
Figure 5

Kurata_etal_FigS1new.pdf (48 kB)
Figure S1

Kurata_etal_FigS2new.pdf (57 kB)
Figure S2

Kurata_etal_FigS3new.pdf (106 kB)
Figure S3

Kurata_etal_FigS4new.pdf (38 kB)
Figure S4

Kurata_etal_FigS5new.pdf (176 kB)
Figure S5

Kurata_etal_FigS6new.pdf (199 kB)
Figure S6

Kurata_etal_FigS7new.pdf (146 kB)
Figure S7

Kurata_etal_FigS8new.pdf (359 kB)
Figure S8

 
 

To view the content in your browser, please download Adobe Reader or, alternately,
you may Download the file to your hard drive.

NOTE: The latest versions of Adobe Reader do not support viewing PDF files within Firefox on Mac OS and if you are using a modern (Intel) Mac, there is no official plugin for viewing PDF files within the browser window.