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. 1969 Oct;4(4):416–422. doi: 10.1128/jvi.4.4.416-422.1969

Synthesis of Vaccinia Viral Proteins in Cytoplasmic Extracts

I. Incorporation of Radioactively Labeled Amino Acids into Polypeptides

Ehud Katz 1, Bernard Moss 1
PMCID: PMC375890  PMID: 5823230

Abstract

Polypeptide synthesis has been studied in cell-free systems prepared from vaccinia virus-infected and uninfected HeLa cells. Cytoplasmic extracts containing endogenous messenger ribonucleic acid were used. Amino acid incorporation into hot trichloroacetic acid-precipitable material was linear for 15 to 20 min at 37 C. The initial rate of protein synthesis was approximately 15% of the rate in intact cells. Optimal conditions for polypeptide synthesis were similar in cell-free systems prepared from infected or uninfected cells. Requirements for an energy source and Mg++ were demonstrated. The optimal Mg++ concentration was 4 to 5 mm. Ribonuclease, puromycin, and cycloheximide were inhibitory. The molecular weights of the polypeptides labeled in the cell-free systems, as determined by gel filtration in 5 m guanidine hydrochloride, ranged from 16,000 to above 68,000. Polyacrylamide gel electrophoresis indicated that the polypeptides labeled in cell-free extracts of uninfected and infected cells were different. The latter closely corresponded in electrophoretic mobility with the viral polypeptides made in intact, infected cells.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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