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Molecular Biology and Gene Regulation

Protein Synthesis in Isolated Mitochondria of Rice (Oryza sativa L.) Seedlings ¹

Institute of Botany, Academia Sinica, Taipei, Taiwan, Republic of China, Taiwan Agriculture Research Institute, Wufeng, Taichung, Taiwan, Republic of China, Department of Ecology and Evolution, The University of Chicago, Chicago, Illinois 60637, Department of Molecular Genetics and Cell Biology, The University of Chicago, Chicago, Illinois 60637

For studies of in organello mitochondrial protein synthesis in rice, Oryza sativa L., conventional surface-sterilization procedures were demonstrated to be ineffective. Because of the over-whelmingly efficient [³⁵S]methionine utilization by contaminating bacteria, even "essentially bacteria-free" rice mitochondria were shown to be unsuitable for the study of in organello protein synthesis. We developed a procedure to obtain a bacteria-free preparation of rice mitochondria. Such mitochondria favored a membrane-dependent ATP-generating system over an external ATP-generating system as the energy supplement for in organello protein synthesis. Two distinct classes of [³⁵S]methionine-labeled, cycloheximide-insensitive products were detected: an electrophoretically unresolved population and a set of some 22 to 27 discrete polypeptide species, each with a characteristic electrophoretic mobility and relative abundance.

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