Induction and de novo synthesis of uricase, a nitrogen-regulated enzyme in Neurospora crassa.
Two efficient procedures are presented for the purification of the purine catabolic enzyme uricase from Neurospora crassa. A specific antiserum for uricase was prepared and used to examine the regulation of uricase expression. Even when wild-type cells are growing under full nitrogen repression conditions, they possess a considerable basal level of uricase. Induction results in a severalfold increase in the level of this enzyme and reflects de novo enzyme synthesis. Identical forms of uricase were translated in vitro from RNA isolated from control and induced cells, but, unexpectedly, induced cells contained less translatable uricase mRNA than did control cells. Although uricase is localized in peroxisomes, the enzyme subunit appears to be synthesized in mature form without any requirement for processing.
D004790:Enzyme Induction; D005656:Fungal Proteins / Q000235:genetics; D005786:Gene Expression Regulation; D005800:Genes, Fungal; D009491:Neurospora / Q000201:enzymology*; D009492:Neurospora crassa / Q000201:enzymology*; D009584:Nitrogen / Q000502:physiology; D014176:Protein Biosynthesis; D011499:Protein Processing, Post-Translational; D012333:RNA, Messenger / Q000235:genetics; D013997:Time Factors; D014503:Urate Oxidase / Q000302:isolation & purification / Q000502:physiology*
D005656:Fungal Proteins; D012333:RNA, Messenger; D014503:Urate Oxidase; D009584:Nitrogen