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A transfer RNA-dependent protein synthesizing system from Ehrlich ascites extracts.
Biochimica Et Biophysica Acta
|May 16, 1975
Summary
Researchers developed a cell-free system dependent on transfer RNA (tRNA) for protein synthesis. Adding ascites cell tRNA significantly boosted protein production with various messenger RNAs (mRNAs).
Area of Science:
- Molecular Biology
- Biochemistry
- Cell Biology
Background:
- Cell-free systems are crucial for studying protein synthesis in vitro.
- Ehrlich ascites cells provide a rich source of cellular components for biochemical assays.
Purpose of the Study:
- To create a transfer RNA (tRNA)-dependent homologous cell-free system for protein synthesis.
- To investigate the role of ascites cell tRNA in stimulating protein production.
Main Methods:
- Developed a homologous cell-free system from Ehrlich ascites cells.
- Rendered the system tRNA-dependent by DEAE-cellulose column chromatography of ribosomes and ammonium sulfate fraction.
- Tested protein synthesis stimulation with varying messenger RNA (mRNA) sources.
Main Results:
- The cell-free system demonstrated robust protein synthesis upon tRNA addition.
- Protein synthesis was enhanced 20-50 fold with the addition of 100 µg/ml ascites cell tRNA.
- Stimulation was observed with oviduct mRNA, ascites mRNA, and encephalomyocarditis RNA.
Conclusions:
- The established cell-free system effectively utilizes ascites cell tRNA to drive protein synthesis.
- This system provides a valuable tool for studying translational regulation and mRNA function.