FGSC #4545

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Reporting Genes: trp-1;qa-2;aro-9;inl

Species: crassa

Allele: 20 M246 Y325M6 89601

Alternate Strain Number: 9313

Depositor: DDP

Linkage Group: IIIR;VII;IIR;VR

Mating Type: A

Species Number: 10

Deposit Sheet ↗

Genes

Locus	Cultural Requirements	Link Group	Type
aro-9	Requires shikimic acid or a mixture of the four aromatic amino acids when combined with qa-1. Single mutant grows on minimal media.	IIR	B
inl	VR. Between pho-3 (3 to 4%) and pab-1 (1 to 10%). Right of al-3 (362, 397, 1036). (482)Requires inositol (65). Lacks D-myoinositol-1-phosphatase (1142). Lack of glucocycloaldolase found by Pina and Tatum (826) is attributed by Williams (1142) to drastic repression of glucocycloaldolase by the concentration of inositol used for growth. Growth is colonial on low levels of inositol (367). Tends to extrude dark pigment into the medium when grown on suboptimal inositol. Composition of phospholipids and cell walls is abnormal on limiting inositol (367, 439, 440, 501). Inhibited by hexachlorocyclohexane (366, 457, 931). Conidia are subject to death by unbalanced growth on minimal medium (1028, 1033), a property exploited for mutant enrichment ("inositol-less death") (606, 647) because double mutants are at a selective advantage. Heat-sensitive allele 83201 is especially useful for mutant enrichment (832, 1043). Used in the first experiments reporting transformation of Neurospora by N. crassaDNA (677, 679) and reported to be efficient as a recipient in absence of inositol (1162). Used to study glucose (917) and sulfate (641) transport systems. Used extensively for studying induced reversion (392). Used for studying the mechanism of inositol-less death (647, 702), mutagenicity of ferrous ions, and regulation of mitochondrial membrane fluidity; for a review, see reference 702. Spontaneous reversion rates (386). Allele-specific partial suppressor (390). Allele 46802 is nonrevertable and inseparable from translocation 46802 (386, 808). Strains carrying heat-sensitive allele 83201 show slow semicolonial growth in liquid minimal medium at 25°C (641), but look normal on slants (D.D. Perkins, unpublished data). Strains carrying allele 89601 contain cross-reacting material (1183). Mutant gene exo-1 is present in the inl(89601) a stock FGSC 498 and may, therefore, be present in stocks of mutants derived by inositol-less death. (See references 194, 325, and 1027). Called inos.	VR	B
qa-2	VII. In the qa cluster between qa-4 and met-7 (0.02%) (146). (884) Lacks catabolic dehydroquinase (148, 517, 884). Unable to use quinate or shikimate as the sole carbon source in the presence of aro-9, which results in the absence of biosynthetic dehydroquinase (885) (Fig. 21). aro-9;qa-2+ strains grow on minimal medium without supplement. qa-2 is conveniently scored as an aro auxotroph when aro-9 is present. qa-2+ cloned in pBR322 (pVK57) is expressed constitutively from its own promoter in E. coli(23, 388, 451); has been returned to Neurospora chromosomal sites by transformation (150). Allele M246 is stable (150).	VII	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