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In vitro synthesis of adenovirus core proteins
Abstract:
mRNA extracted from polysomes of KB cells at late stages of productive infection with adenovirus type 2 was translated in a cell-free system derived from Krebs II ascites cells. Two of the polypeptides obtained in this reaction corresponded to the adenovirus core protein V and the precursor to core protein VII. The following criteria were used to establish identity between the in vitro products and the virion proteins: comigration during sodium dodecyl sulfatepolyacrylamide gel electrophoresis, cochromatography in sodium dodecyl sulfate-hydroxyapatite, specific immunoprecipitation of the precursor to core protein VII, and tryptic peptide analysis.
Insights
Researchers translated adenovirus type 2 mRNA in a cell-free system, identifying core proteins V and VII. This study advances understanding of adenovirus protein synthesis during infection.
Area of Science:
- Molecular Biology
- Virology
- Biochemistry
Background:
- Adenovirus type 2 infection involves complex protein synthesis.
- Understanding viral protein production is crucial for antiviral strategies.
Purpose of the Study:
- To identify adenovirus type 2 core proteins synthesized in vitro.
- To characterize the translation products of late-stage viral mRNA.
Main Methods:
- Cell-free translation of mRNA extracted from infected KB cells.
- Analysis of translation products using SDS-PAGE and SDS-hydroxyapatite chromatography.
- Confirmation of protein identity via immunoprecipitation and tryptic peptide analysis.
Main Results:
- Successfully translated mRNA yielded polypeptides corresponding to adenovirus core protein V.
- Identified a precursor to adenovirus core protein VII among the in vitro products.
- Confirmed protein identities using multiple biochemical and immunological techniques.
Conclusions:
- The cell-free system accurately reflects adenovirus type 2 protein synthesis.
- Identified key viral core proteins, V and a precursor to VII, synthesized during productive infection.