Poly(A) hydrolase of chick-embryo fibroblasts

1. Homogenates of cultured chick embryo fibroblasts have been quantitatively fractionated by differential centrifugation. Using cytochrome c oxidase, succinate dehydrogenase, acid phosphatase and NADPH-cytochrome c reductase as marker enzymes, poly(A) hydrolase has been shown to be a mitochondrial enzyme. 2. To test the biosynthetic origin of mitochondrial poly(A) hydrolase and to demonstrate its cytoplasmic site of synthesis, we have treated the cells with ethidium bromide, inhibitor of mitochondrial transcription, and chloramphenicol and cycloheximide, inhibitors of mitochondrial and cytoplasmic translations respectively. The activity of poly(A) hydrolase has been compared to that of succinate dehydrogenase, an enzyme coded for by the nuclear genome and that of cytochrome c oxidase, an enzyme coded for partly by the nuclear genome and partly by the mitochondrial genome. The results obtained indicate that in chick embryo fibroblasts poly(A) hydrolase is an enzyme coded for by the nuclear genome. Further, the hydrolase is synthesized on cytoplasmic ribosomes and has a half-life much shorter than succinate dehydrogenase and cytochrome c oxidase.