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Originally published as Genetics Published Articles Ahead of Print on May 4, 2007.

Genetics, Vol. 177, 1527-1537, November 2007, Copyright © 2007
doi:10.1534/genetics.107.070771

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Fine-Tuning of Translation Termination Efficiency in Saccharomyces cerevisiae Involves Two Factors in Close Proximity to the Exit Tunnel of the Ribosome

Isabelle Hatin*,{dagger},1, Céline Fabret*,{dagger}, Olivier Namy*,{dagger}, Wayne A. Decatur{ddagger} and Jean-Pierre Rousset*,{dagger}

* IGM, Université Paris-Sud, UMR 8621, F91405 Orsay, France, {dagger} CNRS, F91405 Orsay, France and {ddagger} Department of Biochemistry and Molecular Biology, University of Massachusetts, Amherst, Massachusetts 01003

1 Corresponding author: Institut de Génétique et Microbiologie, Bâtiment 400, Université Paris-Sud, F91405 Orsay, France.
E-mail: isabelle.hatin{at}igmors.u-psud.fr

In eukaryotes, release factors 1 and 3 (eRF1 and eRF3) are recruited to promote translation termination when a stop codon on the mRNA enters at the ribosomal A-site. However, their overexpression increases termination efficiency only moderately, suggesting that other factors might be involved in the termination process. To determine such unknown components, we performed a genetic screen in Saccharomyces cerevisiae that identified genes increasing termination efficiency when overexpressed. For this purpose, we constructed a dedicated reporter strain in which a leaky stop codon is inserted into the chromosomal copy of the ade2 gene. Twenty-five antisuppressor candidates were identified and characterized for their impact on readthrough. Among them, SSB1 and snR18, two factors close to the exit tunnel of the ribosome, directed the strongest antisuppression effects when overexpressed, showing that they may be involved in fine-tuning of the translation termination level.




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