Acyl-Coa oxidase and hydratase-dehydrogenase, two enzymes of the peroxisomal beta-oxidation system, are synthesized on free polysomes of clofibrate-treated rat liver.
We investigated the site of synthesis of two abundant proteins in clofibrate-induced rat hepatic peroxisomes. RNA was extracted from free and membrane-bound polysomes, heated to improve translational efficiency, and translated in the mRNA-dependent, reticulocyte-lysate-cell-free, protein-synthesizing system. The peroxisomal acyl-CoA oxidase and enoyl-CoA hydratase-beta-hydroxyacyl-CoA dehydrogenase 35S-translation products were isolated immunochemically, analyzed by SDS PAGE and fluorography, and quantitated by densitometric scanning. The RNAs coding for these two peroxisomal proteins were found predominantly on free polysomes, and the translation products co-migrated with the mature proteins. As in normal rat liver, preproalbumin and catalase were synthesized mainly by membrane-bound and by free polysomes, respectively. mRNAs for a number of minor 35S-translation products also retained by the anti-peroxisomal immunoadsorbent were similarly found on free polysomes. These results, together with previous data, allow the generalization that the content proteins of rat liver peroxisomes are synthesized on free polysomes, and the data imply a posttranslational packaging mechanism for these major content proteins.
D042962:Acyl-CoA Oxidase; D000818:Animals; D002994:Clofibrate / Q000494:pharmacology*; D004746:Enoyl-CoA Hydratase / Q000096:biosynthesis / Q000235:genetics*; D006836:Hydro-Lyases / Q000235:genetics*; D007700:Kinetics; D008099:Liver / Q000187:drug effects / Q000201:enzymology*; D008297:Male; D008830:Microbodies / Q000187:drug effects / Q000201:enzymology*; D008970:Molecular Weight; D010088:Oxidoreductases / Q000096:biosynthesis / Q000235:genetics*; D011132:Polyribosomes / Q000201:enzymology*; D014176:Protein Biosynthesis / Q000187:drug effects; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D011916:Rats, Inbred F344; D012156:Reticulocytes / Q000378:metabolism
D012333:RNA, Messenger; D010088:Oxidoreductases; D042962:Acyl-CoA Oxidase; D006836:Hydro-Lyases; D004746:Enoyl-CoA Hydratase; D002994:Clofibrate