Abstract
Chinese hamster ovary (CHO) cells were subjected to severe amino acid starvation for histidine, leucine, methionine, asparagine, tyrosine, glutamine, valine, and lysine, using amino acid analogs or mutations in specific aminoacyl-tRNA synthetases. At protein synthetic rates of less than 5%, in all cases, the newly synthesized proteins were found on two- dimensional electrophoretic gels to consist of a few intensely labeled spots, with the exception of lysine. This pattern could also be produced by strong inhibition of cytoplasmic protein synthesis with cycloheximide, and was abolished by preincubation with the mitochondrial protein synthesis inhibitor chloramphenicol. It appears therefore that the spots represent mitochondrial protein synthesis and that animal cells must have separate aminoacyl-tRNA synthetases for mitochondrial tRNAs corresponding to all these amino acids except, possibly, for lysine.
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Selected References
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- Anderson S., Bankier A. T., Barrell B. G., de Bruijn M. H., Coulson A. R., Drouin J., Eperon I. C., Nierlich D. P., Roe B. A., Sanger F. Sequence and organization of the human mitochondrial genome. Nature. 1981 Apr 9;290(5806):457–465. doi: 10.1038/290457a0. [DOI] [PubMed] [Google Scholar]
- Blobel G. Intracellular protein topogenesis. Proc Natl Acad Sci U S A. 1980 Mar;77(3):1496–1500. doi: 10.1073/pnas.77.3.1496. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Harley C. B., Pollard J. W., Chamberlain J. W., Stanners C. P., Goldstein S. Protein synthetic errors do not increase during aging of cultured human fibroblasts. Proc Natl Acad Sci U S A. 1980 Apr;77(4):1885–1889. doi: 10.1073/pnas.77.4.1885. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kolarov J., Kuzela S., Wielburski A., Nelson B. D. The characterization of mitochondrial translation products in rat liver and rat hepatoma cells. FEBS Lett. 1981 Apr 6;126(1):61–65. doi: 10.1016/0014-5793(81)81033-2. [DOI] [PubMed] [Google Scholar]
- O'Farrell P. H. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed] [Google Scholar]
- Parker J., Pollard J. W., Friesen J. D., Stanners C. P. Stuttering: high-level mistranslation in animal and bacterial cells. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1091–1095. doi: 10.1073/pnas.75.3.1091. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pollard J. W., Harley C. B., Chamberlain J. W., Goldstein S., Stanners C. P. Is transformation associated with an increased error frequency in mammalian cells? J Biol Chem. 1982 Jun 10;257(11):5977–5979. [PubMed] [Google Scholar]
- Thompson L. H., Harkins J. L., Stanners C. P. A mammalian cell mutant with a temperature-sensitive leucyl-transfer RNA synthetase. Proc Natl Acad Sci U S A. 1973 Nov;70(11):3094–3098. doi: 10.1073/pnas.70.11.3094. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Thompson L. H., Lofgren D. J., Adair G. M. CHO cell mutants for arginyl-, asparagyl-, glutaminyl-, histidyl- and methionyl-transfer RNA synthetases: identification and initial characterization. Cell. 1977 May;11(1):157–168. doi: 10.1016/0092-8674(77)90326-9. [DOI] [PubMed] [Google Scholar]
- Wallace R. B., Williams T. M., Freeman K. B. Mitochondrial protein synthesis in a mammalian cell-line with a temperature-sensitive leucyl-tRNA synthetase. Eur J Biochem. 1975 Nov 1;59(1):167–173. doi: 10.1111/j.1432-1033.1975.tb02438.x. [DOI] [PubMed] [Google Scholar]