FGSC #1847

Add to Cart

Reporting Genes: trp-1;am

Species: crassa

Allele: 15;am17

Alternate Strain Number: am17.tryp-1(15)

Mutagen: UV

Depositor: JHC

Linkage Group: IIIR;VR

Mating Type: a

Species Number: 10

Genetic Background: SL

Opposite Mating Type: 1848

Deposit Sheet ↗

Genes

Locus	Cultural Requirements	Link Group	Type
am	VR. Right of ure-2 (2%) and sp (4 to 8%). Left of gul-1(<<1%) and ace-5(<1%) (122, 570, 579, 998). (R.W. Barratt, (cited in reference 1036) Structural gene for nicotinamide adenine dinucleotide phosphate (NADP)-glutamate dehydrogenase (336) (see Fig. 19), for which a complete 452-residue amino acid sequence has been obtained (465). Requires a source of alpha-amino nitrogen for growth, alanine being a good supplement (e.g., reference 997). Readily scorable at 25 C; leaky at 34°C (42). Leaky growth and adaptation on minimal medium are prevented by 0.02 M glycine (782, 783) or by en(am)-1, en(am)-2, or nit-2, q.v. The ammutants show abnormal regulation of reduced nicotinamide adenine dinucleotide (NADH)-glutamate dehydrogenase and are synergistic with nit-2 in this effect (226). Some am alleles (e.g., RU1) suppress the pyrimidine requirement caused by pyr-3 (CPS- ACT+) mutations (1137). Used for the first demonstration of complementation between alleles in vivo (344) (simultaneous with independent demonstration with ad-4). In vitro complementation (342). Used for studies of complementation mechanism (199, 200, 1120). Used for fine-structure mapping (337, 338). Control of intralocus recombination by rec-3(996-998). Used to study colinearity of the gene and gene product, internal suppressors (105, 340, 465), and the action of supersuppressors (954, 955). The functional defects in several mutant enzymes with single amino acid replacements have been defined: am1 mutant enzymes fail to bind NADPH (1120); am2, am3, am19, am130, and am131 enzymes are stabilized in the inactive conformational form (30, 200, 336, 556, 1044), and all are complementable by am1; am14 is osmotically reparable and is thought to have unstable quaternary structure (340). Used in a study showing glutamine to have a role as corepressor of uricase synthesis (1118). Used to study nitrogen assimilation and metabolism (503) and nitrogen metabolite repression (186, 291). Efficient procedure for selecting new am mutants (551). Spectrum of ultraviolet irradiation (UV)- and nitrous acid-induced mutants (554). Allele am17 has a chain-terminating codon of either the amber or ochre type at residue 313 of glutamate dehydrogenase, based on amino acid replacements in revertants and by ssu-1 (956). Allele 6 is a frameshift mutation with an insertion in the Ser5 codon (985). Allele 126 is highly unstable (553). Allele 132 is a deletion (1162). The am+ gene has been cloned in Escherichia coli(J.R.S. Fincham, personal communication) and transformed back into Neurospora (J.A. Kinsey, personal communication).	VR	B
trp-1	IIIR. Between ad-2 (1 to 7%) and ro-2 (2 to 12%) (11, 219, 812). Linked to fpr-3 (< 1%) (550). (504). Uses tryptophan or indole (1060); strains carrying some alleles can also use anthranilate; others cannot (4). trp-1+ and trp-2+ gene products together form an enzyme aggregate with three activities: anthranilate synthetase, phosphoribosyl-anthranilate isomerase, and indoleglycerol-phosphate synthetase (181, 260) (Fig. 11). trp-1 codes for the beta subunit of the aggregate (546); it specifies phosphoribosyl-anthranilate isomerase, indoleglycerol-phosphate synthetase, and collaboratively the glutamine amino transferase activity of anthranilate synthetase (29, 181, 502). Strains carrying different alleles differ in lacking one or more of the three activities, e.g., trp-1 (allele 15) lacks all three activities; trp-1 (20) lacks only phosphoribosyl- anthranilate isomerase, trp-1C (1) lacks only anthranilate synthetase, trp-1 (25) lacks both phosphoribosyl-anthranilate isomerase and indoleglycerol-phosphate synthetase, etc. (259). (To avoid confusion, note that in reference 259 and related papers, the same "allele number" may be used for a trp-2 mutation, a trp-1 mutation [non-anthranilate-utilizing], and a trp-1Cmutation [anthranilate utilizing]; mutations of the last class are listed by FGSC as trp-1 with the allele number prefixed by C.) Strains carrying different alleles differ in their ability to form aggregates (181, 259). Association between trp-1 and trp-2products is essential for glutamine-dependent anthranilate synthetase activity but not the other two activities (181). The trp-1 gene has been cloned (545, 925), sequenced (925), and reintroduced into Neurospora by transformation (925). It is only partially expressed in E. coli. Fine-structure maps (10, 259). Complementation maps (10, 163). Reviewed as example of gene fusion (218). Nonsense allele used to demonstrate restoration of normal enzyme aggregate by supersuppressors (183). Alleles that accumulate anthranilate are scorable by blue fluorescence under long-wave UV after 2 to 5 days of growth on minimal medium plus indole (10 µg /ml), 34°C (814, 816). Aging cultures may produce brown pigment; blue fluorescence disappears as pigment forms.	IIIR	B