Decreased accuracy of protein synthesis in extracts from aging human diploid fibroblasts.
The accuracy of protein synthesis has been measured in extracts from human diploid fibroblasts of different ages. Extracts were supplied with purified mRNA for the coat protein of the cowpea variant of tobacco mosaic virus (CcTMV), which lacks codons for cysteine and methionine. The presence of 35S-cysteine in CcTMV coat protein synthesized during translation reactions therefore represents translational error. Translation reactions were performed with extracts from young fibroblasts (less than 50% of life span completed) and old fibroblasts (more than 90% of life span completed), and the translation products were purified by immunoprecipitation and analyzed by polyacrylamide gel electrophoresis. The error frequency increased from 4.2 x 10(-5) cysteines/amino acid in young cell extracts to 2.9 x 10(-4) cysteines/amino acid in old cell extracts. Cysteine incorporation was not due to nonspecific binding, and could be increased approximately sixfold by the addition of the misreading antibiotic, paromomycin. It is concluded that translational accuracy is not stable during aging in vitro, and it is proposed that this decrease in the fidelity of information transfer could be responsible for the variety of changes observed in aging cultured human cells.
Luce
M C
MC
Department of Biology, University of South Carolina, Columbia 29208.
Bunn
C L
CL
D000596:Amino Acids / Q000032:analysis; D002460:Cell Line; D002470:Cell Survival; D003545:Cysteine / Q000378:metabolism; D004171:Diploidy; D004591:Electrophoresis, Polyacrylamide Gel; D005347:Fibroblasts / Q000378:metabolism; D006801:Humans; D011233:Precipitin Tests; D014176:Protein Biosynthesis; D014027:Tobacco Mosaic Virus / Q000235:genetics
D000596:Amino Acids; D003545:Cysteine