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Isolation and characterization of the structural gene encoding elongation factor 3.
M Sandbaken1, J A Lupisella, B DiDomenico
1Medical College of Wisconsin, Milwaukee.
Biochimica Et Biophysica Acta
|August 27, 1990
Summary
The yeast translational factor EF-3 (eukaryotic elongation factor 3) is essential for growth and influences protein synthesis accuracy. Elevated EF-3 levels increase sensitivity to antibiotics that cause translational errors.
Area of Science:
- Molecular Biology
- Yeast Genetics
- Protein Synthesis
Background:
- The eukaryotic elongation factor 3 (EF-3) is a unique translational factor in yeast.
- EF-3 plays a role in the ribosome elongation cycle by enhancing the function of EF-1 alpha in aminoacyl-tRNA binding.
Purpose of the Study:
- To isolate and characterize the structural gene encoding yeast EF-3.
- To investigate the function and potential impact of EF-3 on translational accuracy.
Main Methods:
- Isolation and DNA sequencing of the YEF3 gene from Saccharomyces cerevisiae.
- Analysis of the deduced amino acid sequence for conserved motifs.
- Assessing the effect of YEF3 gene overexpression on ATPase activity and antibiotic sensitivity.
Main Results:
- The YEF3 gene is a single-copy, essential gene for yeast vegetative growth.
- The YEF3 gene encodes a protein with two nucleotide-binding motifs similar to those in ATPases.
- Overexpression of YEF3 leads to increased ribosome-dependent ATPase activity and heightened sensitivity to aminoglycoside antibiotics (hygromycin, paromomycin).
Conclusions:
- Yeast EF-3 is an essential protein involved in translation.
- EF-3's structure suggests a role in ATP hydrolysis.
- Increased EF-3 levels correlate with increased translational errors, implying a role in translational accuracy.