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Translation of endogenous message in embryonic chick erythroid cell polysomes in a cell-free protein-synthesizing
Insights
Chick embryonic erythroid cells contain translatable messenger RNA (mRNA). A wheat germ extract enhances protein synthesis from this mRNA in a cell-free system, suggesting a missing factor.
Area of Science:
- Molecular Biology
- Developmental Biology
- Biochemistry
Background:
- Polysomes from embryonic chick erythroid cells contain messenger RNA (mRNA).
- This mRNA can be translated in cell-free systems.
- Previous studies indicated the presence of globin mRNA in these cells.
Purpose of the Study:
- To characterize the translatable mRNA present in embryonic chick erythroid cell polysomes.
- To investigate factors that may enhance protein synthesis directed by this mRNA.
- To identify the protein products synthesized in a cell-free system.
Main Methods:
- Isolation of polysomes from embryonic chick erythroid cells at 5.5 and 6.5 days of development.
- Cell-free protein synthesis using isolated polysomal mRNA.
- Translation product analysis via co-electrophoresis with carrier globin chains.
- Assessment of protein synthesis enhancement using wheat germ S30 fraction and [3H] leucine incorporation.
Main Results:
- Polysomes from embryonic chick erythroid cells contain mRNA translatable into proteins.
- The synthesized products co-electrophoresed with authentic globin chains.
- Wheat germ S30 fraction stimulated [3H] leucine incorporation by 1.5–2.0 fold, indicating enhanced translation.
Conclusions:
- Embryonic chick erythroid cell mRNA directs the synthesis of globin chains in a cell-free system.
- Wheat germ extract likely provides essential factors missing in the standard incubation medium, enhancing translation efficiency.
Abstract:
Polysomes from 5.5-day and 6.5-day embryonic chick erythroid cells contain messenger RNA (mRNA) which can be translated into products in a cell-free protein-synthesizing system. The products of the cell-free system coelectrophoresed with carrier globin chains from the cells from which the polysomal pellet was isolated. Wheat germ S30 fraction increases by 1.5--2.0 fold [3H] leucine incorporation into trichloroacetic acid-precipitable material directed by the endogenous mRNA on chick erythroid cell polysomes. The wheat germ probably provides a specific factor or factors absent in a shortage in the incubation medium.