The regulation of gene expression is critical for cell growth and development. The success of transcriptional and post-transcriptional mechanisms that regulate gene expression hinge upon the successful synthesis of functional polypeptides. Many soluble translation factors, the aminoacyltRNAs (aa-tRNAs) and the ribosome interact to produce nascent polypeptides chains required by the cell. This proposal uses the translation Elongation Factor eEFIA as a central point from which to examine the interactions between these critical components of the translation machinery that function to ensure accurate and appropriate protein synthesis. The analysis of eEF1A function will dissect the mechanisms by which translational fidelity is ensured and how eEF1A serves to integrate the communication between the aa-tRNA and the ribosome. As a G-protein, work on eEF1A has implications in our understanding of cellular regulatory mechanism through nucleotide exchange and hydrolysis. The yeast Saccharomyces cerevisiae allows a combined genetic and biochemical approach to address key questions regarding the mechanism of elongation and the maintenance of translational accuracy. While these studies have implications for all eukaryotic systems, the interaction of eEF1A with the fungal specific translation factor eEF3 defines a process unique to fungi. The interaction of eEF3 with eEF1A and the ribosome, and its essential function in vivo, support the role of eEF3 as a key player in fungal protein synthesis. Roles for eEF3 in translational accuracy have also been proposed, however, little is known about the effects of eEF3 on fidelity in vivo. Accurate protein synthesis is a critical step in ensuring the regulatory mechanisms that exist for modulating gene expression are productive. Interestingly, the accuracy of translation elongation is a process many viruses manipulate to complete a critical part of their life cycle. The proposed studies address the critical communication involved in ensuring translational accuracy in eukaryotes, while potentially yielding new insights into viral-host interactions. Additionally, the analysis of a fungal specific translation factor enhances our understanding of a unique aspect of protein synthesis and may yield new anti-fungal targets.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM057483-09
Application #
7105032
Study Section
Physiological Chemistry Study Section (PC)
Program Officer
Rhoades, Marcus M
Project Start
1998-09-01
Project End
2007-12-06
Budget Start
2006-08-01
Budget End
2007-12-06
Support Year
9
Fiscal Year
2006
Total Cost
$359,231
Indirect Cost
Name
University of Medicine & Dentistry of NJ
Department
Genetics
Type
Schools of Medicine
DUNS #
617022384
City
Piscataway
State
NJ
Country
United States
Zip Code
08854
Mateyak, Maria K; Kinzy, Terri Goss (2017) Breaking the Silos of Protein Synthesis. Trends Biochem Sci 42:587-588
Dever, Thomas E; Kinzy, Terri Goss; Pavitt, Graham D (2016) Mechanism and Regulation of Protein Synthesis in Saccharomyces cerevisiae. Genetics 203:65-107
Sasikumar, Arjun N; Kinzy, Terri Goss (2014) Mutations in the chromodomain-like insertion of translation elongation factor 3 compromise protein synthesis through reduced ATPase activity. J Biol Chem 289:4853-60
Li, Zhenghe; Gonzalez, Paulina Alatriste; Sasvari, Zsuzsanna et al. (2014) Methylation of translation elongation factor 1A by the METTL10-like See1 methyltransferase facilitates tombusvirus replication in yeast and plants. Virology 448:43-54
Perez, Winder B; Kinzy, Terri Goss (2014) Translation elongation factor 1A mutants with altered actin bundling activity show reduced aminoacyl-tRNA binding and alter initiation via eIF2? phosphorylation. J Biol Chem 289:20928-38
Sasikumar, Arjun N; Perez, Winder B; Kinzy, Terri Goss (2012) The many roles of the eukaryotic elongation factor 1 complex. Wiley Interdiscip Rev RNA 3:543-55
Sasvari, Zsuzsanna; Izotova, Lara; Kinzy, Terri Goss et al. (2011) Synergistic roles of eukaryotic translation elongation factors 1Býý and 1A in stimulation of tombusvirus minus-strand synthesis. PLoS Pathog 7:e1002438
Mateyak, Maria K; Kinzy, Terri Goss (2010) eEF1A: thinking outside the ribosome. J Biol Chem 285:21209-13
Li, Zhenghe; Pogany, Judit; Tupman, Steven et al. (2010) Translation elongation factor 1A facilitates the assembly of the tombusvirus replicase and stimulates minus-strand synthesis. PLoS Pathog 6:e1001175
Esposito, Anthony M; Mateyak, Maria; He, Dongming et al. (2010) Eukaryotic polyribosome profile analysis. J Vis Exp :

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