The products of mitochondria-bound cytoplasmic polysomes in yeast.
Experiments were undertaken to examine the fate and composition of polypeptides synthesized on cytoplasmic polysomes associated with the outer mitochondrial membrane of Saccharomyces cerevisiae. Mitochondria with their associated cytoplasmic polysomes were isolated from growing yeast spheroplasts and placed in a polypeptide chain completion system together with [35S]methionine. Of the total products synthesized in the readout system, 80 to 85% remain associated with the mitochondria after sucrose gradient centrifugation. Most of the labeled products are resistant to papain digestion unless the membranes are disrupted by treatment with detergent or shaking with glass beads. When free cytoplasmic polysomes were translated in the presence of [35S]methionine and incubated with mitochondria, only about 20% of the labeled polypeptides remain associated with the mitochondria; furthermore, most of these products are equally sensitive to papain digestion in the presence or absence of detergent. These results support the view that the cytoplasmic polysomes associated with the outer mitochondrial membrane of yeast facilitate the segregation of newly synthesized proteins into the organelle. The proportion of the alpha, beta, and gamma subunits of the F1-ATPase was determined among the products synthesized by mitochondria-bound and free cytoplasmic polysomes. By double antibody precipitation and immunoreplicate electrophoresis, we find that the proportion of the subunits of F1-ATPase is much greater among the products of the mitochondria-bound polysomes than those synthesized on free polysomes.
D000251:Adenosine Triphosphatases / Q000096:biosynthesis; D005656:Fungal Proteins / Q000096:biosynthesis*; D007425:Intracellular Membranes / Q000378:metabolism; D007700:Kinetics; D008928:Mitochondria / Q000378:metabolism*; D010206:Papain; D011132:Polyribosomes / Q000378:metabolism*; D014176:Protein Biosynthesis; D006180:Proton-Translocating ATPases; D012441:Saccharomyces cerevisiae / Q000378:metabolism*
D005656:Fungal Proteins; D010206:Papain; D000251:Adenosine Triphosphatases; D006180:Proton-Translocating ATPases