Abstract
The regulation of the glutamate dehydrogenases was investigated in wild-type Neurospora crassa and two classes of mutants altered in the assimilation of inorganic nitrogen, as either nitrate or ammonium. In the wild-type strain, a high nutrient carbon concentration increased the activity of reduced nicotinamide adenine dinucleotide phosphate (NADPH)-glutamate dehydrogenase and decreased the activity of reduced nicotinamide adenine dinucleotide (NADH)-glutamate dehydrogenase. A high nutrient nitrogen concentration had the opposite effect, increasing NADH-glutamate dehydrogenase and decreasing NADPH-glutamate dehydrogenase. The nit-2 mutants, defective in many nitrogen-utilizing enzymes and transport systems, exhibited low enzyme activities after growth on a high sucrose concentration: NADPH-glutamate dehydrogenase activity was reduced 4-fold on NH4Cl medium, and NADH-glutamate dehydrogenase, 20-fold on urea medium. Unlike the other affected enzymes of nit-2, which are present only in basal levels, the NADH-glutamate dehydrogenase activity was found to be moderately enhanced when cells were grown on a low carbon concentration. This finding suggests that the control of this enzyme in nit-2 is hypersensitive to catabolite repression. The am mutants, which lack NADPH-glutamate dehydrogenase activity, possessed basal levels of NADH-glutamate dehydrogenase activity after growth on urea or l-aspartic acid media, like the wild-type strain, and possessed moderate levels (although three- to fourfold lower than the wild-type strain) on l-asparagine medium or l-aspartic acid medium containing NH4Cl. These regulatory patterns are identical to those of the nit-2 mutants. Thus, the two classes of mutants exhibit a common defect in NADH-glutamate dehydrogenase regulation. Double mutants of nit-2 and am had lower NADH-glutamate dehydrogenase activities than either parent. A carbon metabolite is proposed to be the repressor of NADH-glutamate dehydrogenase in N. crassa.
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- Antoine A. D. Purification and properties of the nitrate reductase isolated from Neurospora crassa mutant nit-3. Kinetics, molecular weight determination, and cytochrome involvement. Biochemistry. 1974 May 21;13(11):2289–2294. doi: 10.1021/bi00708a008. [DOI] [PubMed] [Google Scholar]
- Arst H. N., Jr, Cove D. J. Nitrogen metabolite repression in Aspergillus nidulans. Mol Gen Genet. 1973 Nov 2;126(2):111–141. doi: 10.1007/BF00330988. [DOI] [PubMed] [Google Scholar]
- Blumenthal K. M., Smith E. L. Nicotinamide adenine dinucleotide phosphate-specific glutamate dehydrogenase of Neurospora. I. Isolation, subunits, amino acid composition, sulfhydryl groups, and identification of a lysine residue reactive with pyridoxal phosphate and N-ethylmaleimide. J Biol Chem. 1973 Sep 10;248(17):6002–6008. [PubMed] [Google Scholar]
- Brett M., Chambers G. K., Holder A. A., Fincham J. R., Wootton J. C. Mutational amino acid replacements in Neurospora crassa NADP-specific glutamate dehydrogenase. J Mol Biol. 1976 Sep 5;106(1):1–22. doi: 10.1016/0022-2836(76)90297-7. [DOI] [PubMed] [Google Scholar]
- Chang H. C., Mulkins G. J., Dyer J. C., Sorger G. J. Enzymatic and non-enzymatic reduction of nitrite by extracts of Neurospora crassa. J Bacteriol. 1975 Aug;123(2):755–758. doi: 10.1128/jb.123.2.755-758.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chang H. C., Sorger G. J. Effect of ammonium ions on the induction of nitrite reductase in Neurospora crassa. J Bacteriol. 1976 May;126(2):1002–1004. doi: 10.1128/jb.126.2.1002-1004.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dantzig A. H., Zurowski W. K., Ball T. M., Nason A. Induction and repression of nitrate reductase in Neurospora crassa. J Bacteriol. 1978 Feb;133(2):671–679. doi: 10.1128/jb.133.2.671-679.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dubois E., Grenson M., Wiame J. M. Release of the "ammonia effect" on three catabolic enzymes by NADP-specific glutamate dehydrogenaseless mutations in Saccharomyces cerevisiae. Biochem Biophys Res Commun. 1973 Feb 20;50(4):967–972. doi: 10.1016/0006-291x(73)91500-3. [DOI] [PubMed] [Google Scholar]
- FINCHAM J. R. Genetically determined multiple forms of glutamic dehydrogenase in Neurospora crassa. J Mol Biol. 1962 Apr;4:257–274. doi: 10.1016/s0022-2836(62)80004-7. [DOI] [PubMed] [Google Scholar]
- Garrett R. H., Nason A. Further purification and properties of Neurospora nitrate reductase. J Biol Chem. 1969 Jun 10;244(11):2870–2882. [PubMed] [Google Scholar]
- Garrett R. H., Nason A. Involvement of a B-type cytochrome in the assimilatory nitrate reductase of Neurospora crassa. Proc Natl Acad Sci U S A. 1967 Oct;58(4):1603–1610. doi: 10.1073/pnas.58.4.1603. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Grenson M., Hou C. Ammonia inhibition of the general amino acid permease and its suppression in NADPH-specific glutamate dehydrogenaseless mutants of saccharomyces cerevisiae. Biochem Biophys Res Commun. 1972 Aug 21;48(4):749–756. doi: 10.1016/0006-291x(72)90670-5. [DOI] [PubMed] [Google Scholar]
- Hanson M. A., Marzluf G. A. Control of the synthesis of a single enzyme by multiple regulatory circuits in Neurospora crassa. Proc Natl Acad Sci U S A. 1975 Apr;72(4):1240–1244. doi: 10.1073/pnas.72.4.1240. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hynes M. J. Effects of ammonium, L-glutamate, and L-glutamine on nitrogen catabolism in Aspergillus nidulans. J Bacteriol. 1974 Dec;120(3):1116–1123. doi: 10.1128/jb.120.3.1116-1123.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kapoor M., Grover A. K. Catabolite-controlled regulation of glutamate dehydrogenases of Neurospora crassa. Can J Microbiol. 1970 Jan;16(1):33–40. doi: 10.1139/m70-006. [DOI] [PubMed] [Google Scholar]
- Kinghorn J. R., Pateman J. A. NAD and NADP l-glutamate dehydrogenase activity and ammonium regulation in Aspergillus nidulans. J Gen Microbiol. 1973 Sep;78(1):39–46. doi: 10.1099/00221287-78-1-39. [DOI] [PubMed] [Google Scholar]
- LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
- Lafferty M. A., Garrett R. H. Purification and properties of the Neurospora crassa assimilatory nitrite reductase. J Biol Chem. 1974 Dec 10;249(23):7555–7567. [PubMed] [Google Scholar]
- NASON A., ABRAHAM R. G., AVERBACH B. C. The enzymic reduction of nitrite to ammonia by reduced pyridine nucleotides. Biochim Biophys Acta. 1954 Sep;15(1):159–161. doi: 10.1016/0006-3002(54)90118-3. [DOI] [PubMed] [Google Scholar]
- NASON A., EVANS H. J. Triphosphopyridine nucleotide-nitrate reductase in Neurospora. J Biol Chem. 1953 Jun;202(2):655–673. [PubMed] [Google Scholar]
- Nason A., Antoine A. D., Ketchum P. A., Frazier W. A., 3rd, Lee D. K. Formation of assimilatory nitrate reductase by in vitro inter-cistronic complementation in Neurospora crassa. Proc Natl Acad Sci U S A. 1970 Jan;65(1):137–144. doi: 10.1073/pnas.65.1.137. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pan S. S., Nason A. Purification and characterization of homogeneous assimilatory reduced nicotinamide adenine dinucleotide phosphate-nitrate reductase from Neurospora crassa. Biochim Biophys Acta. 1978 Apr 12;523(2):297–313. doi: 10.1016/0005-2744(78)90033-5. [DOI] [PubMed] [Google Scholar]
- Pateman J. A. Regulation of synthesis of glutamate dehydrogenase and glutamine synthetase in micro-organisms. Biochem J. 1969 Dec;115(4):769–775. doi: 10.1042/bj1150769. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pendyala L., Wellman A. M. Purine base transport in nit-2 mutants of Neurospora crassa. J Bacteriol. 1978 Jan;133(1):401–402. doi: 10.1128/jb.133.1.401-402.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- SANWAL B. D., LATA M. Concurrent regulation of glutamic acid dehydrogenases of Neurospora. Arch Biochem Biophys. 1962 Jun;97:582–588. doi: 10.1016/0003-9861(62)90127-3. [DOI] [PubMed] [Google Scholar]
- SANWAL B. D., LATA M. Effect of glutamic acid on the formation of two glutamic acid dehydrogenases of Neurospora. Biochem Biophys Res Commun. 1962 Jan 24;6:404–409. doi: 10.1016/0006-291x(62)90364-9. [DOI] [PubMed] [Google Scholar]
- SANWAL B. D., LATA M. The occurrence of two different glutamic dehydrogenases in Neurospora. Can J Microbiol. 1961 Jun;7:319–328. doi: 10.1139/m61-039. [DOI] [PubMed] [Google Scholar]
- SANWAL B. D., LATA M. The regulation of glutamic dehydrogenases and an antigenically related protein in amination deficient mutants of Neurospora. Arch Biochem Biophys. 1962 Sep;98:420–426. doi: 10.1016/0003-9861(62)90207-2. [DOI] [PubMed] [Google Scholar]
- SILVER W. S., McELROY W. D. Enzyme studies on nitrate and nitrite mutants of Neurospora. Arch Biochem Biophys. 1954 Aug;51(2):379–394. doi: 10.1016/0003-9861(54)90493-2. [DOI] [PubMed] [Google Scholar]
- Schloemen R. H., Garrett R. H. Nitrate transport system in Neurospora crassa. J Bacteriol. 1974 Apr;118(1):259–269. doi: 10.1128/jb.118.1.259-269.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sorger G. J., Giles N. H. Genetic control of nitrate reductase in Neurospora crassa. Genetics. 1965 Oct;52(4):777–788. doi: 10.1093/genetics/52.4.777. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Stachow C. S., Sanwal B. D. Regulation, purification, and some properties of the NAD-specific glutamate dehydrogenase of Neurospora. Biochim Biophys Acta. 1967 Jul 11;139(2):294–307. doi: 10.1016/0005-2744(67)90033-2. [DOI] [PubMed] [Google Scholar]
- Tsao T. F., Marzluf G. A. Genetic and metabolic regulation of purine base transport in Neurospora crassa. Mol Gen Genet. 1976 Dec 22;149(3):347–355. doi: 10.1007/BF00268537. [DOI] [PubMed] [Google Scholar]
- Veronese F. M., Nyc J. F., Degani Y., Brown D. M., Smith E. L. Nicotinamide adenine dinucleotide-specific glutamate dehydrogenase of Neurospora. I. Purification and molecular properties. J Biol Chem. 1974 Dec 25;249(24):7922–7928. [PubMed] [Google Scholar]
- Wootton J. C., Chambers G. K., Taylor J. G., Fincham J. R. Amino-acid sequence homologies between the NADP-dependent glutamate dehydrogenase of Neurospora and the bovine enzyme. Nat New Biol. 1973 Jan 10;241(106):42–43. doi: 10.1038/newbio241042a0. [DOI] [PubMed] [Google Scholar]