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. 1987 May;169(5):1943–1948. doi: 10.1128/jb.169.5.1943-1948.1987

Induction and de novo synthesis of uricase, a nitrogen-regulated enzyme in Neurospora crassa.

B H Nahm, G A Marzluf
PMCID: PMC212054  PMID: 2952636

Abstract

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.

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Selected References

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  1. Blobel G. Intracellular protein topogenesis. Proc Natl Acad Sci U S A. 1980 Mar;77(3):1496–1500. doi: 10.1073/pnas.77.3.1496. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Dunn-Coleman N. S., Garrett R. H. The role fo glutamine synthetase and glutamine metabolism in nitrogen metabolite repression, a regulatory phenomenon in the lower eukaryote Neurospora crassa. Mol Gen Genet. 1980;179(1):25–32. doi: 10.1007/BF00268442. [DOI] [PubMed] [Google Scholar]
  3. Goldman B. M., Blobel G. Biogenesis of peroxisomes: intracellular site of synthesis of catalase and uricase. Proc Natl Acad Sci U S A. 1978 Oct;75(10):5066–5070. doi: 10.1073/pnas.75.10.5066. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Grove G., Marzluf G. A. Identification of the product of the major regulatory gene of the nitrogen control circuit of Neurospora crassa as a nuclear DNA-binding protein. J Biol Chem. 1981 Jan 10;256(1):463–470. [PubMed] [Google Scholar]
  5. Kessler S. W. Cell membrane antigen isolation with the staphylococcal protein A-antibody adsorbent. J Immunol. 1976 Nov;117(5 Pt 1):1482–1490. [PubMed] [Google Scholar]
  6. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  7. Lyon E. S., Garrett R. H. Regulation, purification, and properties of xanthine dehydrogenase in Neurospora crassa. J Biol Chem. 1978 Apr 25;253(8):2604–2614. [PubMed] [Google Scholar]
  8. Marzluf G. A. Regulation of nitrogen metabolism and gene expression in fungi. Microbiol Rev. 1981 Sep;45(3):437–461. doi: 10.1128/mr.45.3.437-461.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Perrine K. G., Marzluf G. A. Amber nonsense mutations in regulatory and structural genes of the nitrogen control circuit of Neurospora crassa. Curr Genet. 1986;10(9):677–684. doi: 10.1007/BF00410916. [DOI] [PubMed] [Google Scholar]
  10. Premakumar R., Sorger G. J., Gooden D. Nitrogen metabolite repression of nitrate reductase in Neurospora crassa. J Bacteriol. 1979 Mar;137(3):1119–1126. doi: 10.1128/jb.137.3.1119-1126.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Reinert W. R., Marzluf G. A. Genetic and metabolic control of the purine catabolic enzymes of Neurospora crasse. Mol Gen Genet. 1975 Aug 5;139(1):39–55. doi: 10.1007/BF00267994. [DOI] [PubMed] [Google Scholar]
  12. Reinert W. R., Marzluf G. A. Regulation of the purine catabolic enzymes in Neurospora crassa. Arch Biochem Biophys. 1975 Feb;166(2):565–574. doi: 10.1016/0003-9861(75)90421-x. [DOI] [PubMed] [Google Scholar]
  13. Sikora L. A., Marzluf G. A. Regulation of L-phenylalanine ammonia-lyase by L-phenylalanine and nitrogen in Neurospora crassa. J Bacteriol. 1982 Jun;150(3):1287–1291. doi: 10.1128/jb.150.3.1287-1291.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Sikora L., Marzluf G. A. Regulation of L-amino acid oxidase and of D-amino acid oxidase in Neurospora crassa. Mol Gen Genet. 1982;186(1):33–39. doi: 10.1007/BF00422908. [DOI] [PubMed] [Google Scholar]
  15. Tolbert N. E. Metabolic pathways in peroxisomes and glyoxysomes. Annu Rev Biochem. 1981;50:133–157. doi: 10.1146/annurev.bi.50.070181.001025. [DOI] [PubMed] [Google Scholar]
  16. Walter P., Gilmore R., Blobel G. Protein translocation across the endoplasmic reticulum. Cell. 1984 Aug;38(1):5–8. doi: 10.1016/0092-8674(84)90520-8. [DOI] [PubMed] [Google Scholar]
  17. Wang L. W., Marzluf G. A. Nitrogen regulation of uricase synthesis in Neurospora crassa. Mol Gen Genet. 1979 Nov;176(3):385–392. doi: 10.1007/BF00333102. [DOI] [PubMed] [Google Scholar]
  18. Wang L. W., Marzluf G. A. Purification and characterization of uricase, a nitrogen-regulated enzyme, from Neurospora crassa. Arch Biochem Biophys. 1980 Apr 15;201(1):185–193. doi: 10.1016/0003-9861(80)90501-9. [DOI] [PubMed] [Google Scholar]

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