Abstract
Cell-free protein synthesis was studied in cytoplasmic S-30 supernatant fractions from uninfected KB cells and from KB cells 18 hr after infection with adenovirus type 2 (Ad 2). Optimal conditions for protein synthesis, including Mg 2+ concentration, energy source, and sulfhydryl compounds, are similar for S-30 fractions prepared from uninfected and infected cells. Amino acid incorporation into trichloracetic acid-precipitable material is linear for 10–15 min at 37° and is inhibited 90–95% by RNase, puromycin, or cycloheximide and 30–40% by sodium fluoride. Approximately 30–40% of the proteins synthesized during 20 min of incorporation are released from ribosomes. Ribosomes in the S-30 fraction from infected cells are programmed by endogenous viral mRNA and synthesize and release all known virion polypeptides in vitro including hexon, penton, fiber, and internal core polypeptides. Double-label coelectrophoresis on polyacrylamide gels showed that the same viral polypeptides are synthesized in vivo and in vitro.
Published Version
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