Chloramphenicol-sensitive mitochondrial translation in Trypanosoma brucei
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- Nabholz, C., Speijer, D. & Schneider, A. Parasitol Res (1999) 85: 779. doi:10.1007/s004360050631
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We developed an in organello system to label newly synthesized mitochondrially encoded proteins of Trypanosoma brucei. Highly purified mitochondria, prepared under isotonic conditions, were incubated with radioactive methionine and cysteine in a suitable translation buffer. Analysis of mitochondrial extracts on TRIS-Tricine gels revealed a subset of labeled, NP-40-insoluble proteins. The labeling of these proteins was resistant to the cytosol-specific translation inhibitor cycloheximide. The proteins, however, were not labeled in the presence of chloramphenicol or erythromycin, inhibitors of prokaryotic type translation, or puromycin, a general translation inhibitor. These results indicate that isotonically isolated mitochondria of T. brucei are capable of protein synthesis.