The overall objective of the proposed research is to elucidate the mechanism of protein biosynthesis in the mitochondria of animal cells and to compare this system to those of prokaryotes and the cytoplasm of eukaryotes. Our initial goal will be to establish an in vitro system from bovine liver mitochondria that will translate both synthetic and natural messenger RNAs. We will then seek to isolate, purify and characterize the mitochondrial factors required for the translation process in this organelle. Our initial efforts will be directed toward the isolation of the elongation factors. The roles of these factors in the translation process will be defined and the nature of the elongation cycle in animal mitochondria will be established. The properties of the purified factors will be examined in detail and they will be compared to the elongation factors of prokaryotes and of the eukaryotic cell cytoplasm. Our second objective will be to purify the initiation factors, describe their properties and define their roles in the initiation process. In addition, we will seek to determine how the initiation signal is recognized on mitochondrial messenger RNA. The results obtained will permit us to compare the process of protein chain initiation in this cell organelle to that in bacteria and in the cell cytoplasm. A long-term objective is to develop a defined in vitro protein synthesizing system from animal cell mitochondria in which to study the regulation of organelle protein synthesis and its integration into the complex metabolism of the eukaryotic cell.
Showing the most recent 10 out of 70 publications
