scholarly journals Dominant-negative mutant phenotypes and the regulation of translation elongation factor 2 levels in yeast

2005 ◽  
Vol 33 (18) ◽  
pp. 5740-5748 ◽  
Author(s):  
P. A. Ortiz
Keyword(s):  
Elongation Factor ◽  
Dominant Negative ◽  
Dominant Negative Mutant ◽  
Translation Elongation Factor ◽  
Translation Elongation ◽  
Elongation Factor 2 ◽  
Regulation Of Translation ◽  
Mutant Phenotypes ◽  
Negative Mutant

scholarly journals Translation elongation factor 2 depletion by siRNA in mouse liver leads to mTOR-independent translational upregulation of ribosomal protein genes

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Maxim V. Gerashchenko ◽  
Mikhail V. Nesterchuk ◽  
Elena M. Smekalova ◽  
Joao A. Paulo ◽  
Piotr S. Kowalski ◽  
...  
Keyword(s):  
Protein Synthesis ◽  
Mammalian Cells ◽  
Ribosomal Proteins ◽  
Mouse Liver ◽  
Negative Impact ◽  
Elongation Factor ◽  
Ribosome Profiling ◽  
Translation Elongation Factor ◽  
Translation Elongation ◽  
Elongation Factor 2

Abstract Due to breakthroughs in RNAi and genome editing methods in the past decade, it is now easier than ever to study fine details of protein synthesis in animal models. However, most of our understanding of translation comes from unicellular organisms and cultured mammalian cells. In this study, we demonstrate the feasibility of perturbing protein synthesis in a mouse liver by targeting translation elongation factor 2 (eEF2) with RNAi. We were able to achieve over 90% knockdown efficacy and maintain it for 2 weeks effectively slowing down the rate of translation elongation. As the total protein yield declined, both proteomics and ribosome profiling assays showed robust translational upregulation of ribosomal proteins relative to other proteins. Although all these genes bear the TOP regulatory motif, the branch of the mTOR pathway responsible for translation regulation was not activated. Paradoxically, coordinated translational upregulation of ribosomal proteins only occurred in the liver but not in murine cell culture. Thus, the upregulation of ribosomal transcripts likely occurred via passive mTOR-independent mechanisms. Impaired elongation sequesters ribosomes on mRNA and creates a shortage of free ribosomes. This leads to preferential translation of transcripts with high initiation rates such as ribosomal proteins. Furthermore, severe eEF2 shortage reduces the negative impact of positively charged amino acids frequent in ribosomal proteins on ribosome progression.

scholarly journals A Role for the Ppz Ser/Thr Protein Phosphatases in the Regulation of Translation Elongation Factor 1Bα

2001 ◽  
Vol 276 (18) ◽  
pp. 14829-14834 ◽  
Author(s):  
Eulàlia de Nadal ◽  
Robert P. Fadden ◽  
Amparo Ruiz ◽  
Timothy Haystead ◽  
Joaquı́n Ariño
Keyword(s):  
Protein Phosphatases ◽  
Elongation Factor ◽  
Translation Elongation Factor ◽  
Translation Elongation ◽  
Regulation Of Translation

scholarly journals A Chemical Genomic Screen in Saccharomyces cerevisiae Reveals a Role for Diphthamidation of Translation Elongation Factor 2 in Inhibition of Protein Synthesis by Sordarin

2008 ◽  
Vol 52 (5) ◽  
pp. 1623-1629 ◽  
Author(s):  
Javier Botet ◽  
María Rodríguez-Mateos ◽  
Juan P. G. Ballesta ◽  
José Luis Revuelta ◽  
Miguel Remacha
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Protein Synthesis ◽  
Elongation Factor ◽  
Translation Elongation Factor ◽  
Cellular Mechanisms ◽  
Translation Elongation ◽  
Chemical Genomic ◽  
Elongation Factor 2 ◽  
Fungal Organisms ◽  
Inhibitor Of Protein Synthesis

ABSTRACT Sordarin and its derivatives are antifungal compounds of potential clinical interest. Despite the highly conserved nature of the fungal and mammalian protein synthesis machineries, sordarin is a selective inhibitor of protein synthesis in fungal organisms. In cells sensitive to sordarin, its mode of action is through preventing the release of translation elongation factor 2 (eEF2) during the translocation step, thus blocking protein synthesis. To further investigate the cellular components required for the effects of sordarin in fungal cells, we have used the haploid deletion collection of Saccharomyces cerevisiae to systematically identify genes whose deletion confers sensitivity or resistance to the compound. Our results indicate that genes in a number of cellular pathways previously unknown to play a role in sordarin response are involved in its growth effects on fungal cells and reveal a specific requirement for the diphthamidation pathway of cells in causing eEF2 to be sensitive to the effects of sordarin on protein synthesis. Our results underscore the importance of the powerful genomic tools developed in yeast (Saccharomyces cerevisiae) to more comprehensively understanding the cellular mechanisms involved in the response to therapeutic agents.

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