Nitrate reactive structural gene mutants of Aspergillus nidulans
Downey, R.J.; Gagliardi, R.J.
Microbios 66(267): 107-116
1991
ISSN/ISBN: 0026-2633 PMID: 2062247 Document Number: 386388
Two strains characterized as niaD structural gene mutants in Aspergillus nidulans produce a nitrate reductase which retains the ability to react with nitrate while lacking the ability to oxidize its naturally occurring substrate NADPH. Fifteen such nitrate reactive niaD strains exhibited strong interallelic complementation when tested against strains bearing point mutations in eleven other loci essential to induction and synthesis of nitrate reductase in Aspergillus. Fourteen representatives of this phenotype formed enzyme with a molecular weight equivalent to that of the wild type (200 kD) and also remained inducible by nitrate and repressible by ammonium. The mutation appears to alter the NADPH binding domain of the nitrate reductase since the affinity for the dinucleotide fold in Affigel blue and the dissociation constant (Ks) for enzyme isolated from the mutants on the basis of reduced methyl viologen-nitrate reductase activity is significantly less than that observed for the native enzyme from the wild type.