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Nucleic Acids Research 2005 33(14):4584-4592; doi:10.1093/nar/gki765
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Published online 12 August 2005

© The Author 2005. Published by Oxford University Press. All rights reserved
The online version of this article has been published under an open access model. Users are entitled to use, reproduce, disseminate, or display the open access version of this article for non-commercial purposes provided that: the original authorship is properly and fully attributed; the Journal and Oxford University Press are attributed as the original place of publication with the correct citation details given; if an article is subsequently reproduced or disseminated not in its entirety but only in part or as a derivative work this must be clearly indicated. For commercial re-use, please contact journals.permissions{at}oupjournals.org

Article

Interplay between GCN2 and GCN4 expression, translation elongation factor 1 mutations and translational fidelity in yeast

Tanya Magazinnik1, Monika Anand1, Evelyn Sattlegger2, Alan G. Hinnebusch2 and Terri Goss Kinzy1,3,*

1Department of Molecular Genetics, Microbiology and Immunology, UMDNJ Robert Wood Johnson Medical School Piscataway, NJ 08854, USA 2Laboratory of Gene Regulation and Development, NICHD, National Institutes of Health Bethesda, MD 20892, USA 3The Cancer Institute of New Jersey, NICHD, National Institutes of Health Bethesda, MD 20892, USA

*To whom correspondence should be addressed. Tel: +1 732 235 5450; Fax: +1 732 235 5223; Email: kinzytg{at}umdnj.edu

Received July 1, 2005. Revised July 26, 2005. Accepted July 26, 2005.

Genetic screens in Saccharomyces cerevisiae have identified the roles of ribosome components, tRNAs and translation factors in translational fidelity. These screens rely on the suppression of altered start codons, nonsense codons or frameshift mutations in genes involved in amino acid or nucleotide metabolism. Many of these genes are regulated by the General Amino Acid Control (GAAC) pathway. Upon amino acid starvation, the kinase GCN2 induces the GAAC cascade via increased translation of the transcriptional activator GCN4 controlled by upstream open reading frames (uORFs). Overexpression of the GCN2 or GCN4 genes enhances the sensitivity of translation fidelity assays that utilize genes regulated by GCN4, such as the suppression of a +1 insertion by S.cerevisiae translation elongation factor 1A (eEF1A) mutants. Paromomycin and the prion [PSI+], which reduce translational fidelity, do not increase GCN4 expression to induce the suppression phenotype and in fact reduce derepression. eEF1A mutations that reduce translation, however, reduce expression of GCN4 under non-starvation conditions. These eEF1A mutants also reduce HIS4 mRNA expression. Taken together, this system improves in vivo strategies for the analysis of translational fidelity and further provides new information on the interplay among translation fidelity, altered elongation and translational control via uORFs.

Present address: Evelyn Sattlegger, Institute of Molecular BioSciences, Massey University, Private Bag 102 904 NSMC, Auckland, New Zealand


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