Effect of mutations in the qa gene cluster of Neurospora crassa on the enzyme catabolic dehydroquinase.
Catabolic dehydroquinase, which functions in the inducible quinic acid catabolic pathway of Neurospora crassa, has been purified from wild type (74-A) and three mutants in the qa gene cluster. The mutant strains were: 105c, a temperature-sensitive constitutive mutant in the qa-1 regulatory locus; M-16, a qa-3 mutant deficient in quinate dehydrogenase activity; and 237, a leaky qa-2 mutant which possess very low levels of catabolic dehydroquinase activity. The enzymes purified from strains 74-A, 105c, and M-16 are identical with respect to behavior during purification, specific activity, electrophoretic behavior, stability, molecular weight, subunit structure, immunological cross-reactivity, and amino acid content. The mutant enzyme from strain 237 is 1,500-fold less active and appears to have a slightly different amino acid content. It is identical by a number of the other criteria listed above and is presumed to be a mutant at or near the enzyme active site. These data demonstrate that the qa-1 gene product is not involved in the posttranslational expression of enzyme activity. The biochemical identity of catabolic dehydroquinase isolated from strains 105c and M-16 with that from wild type also demonstrates that neither the inducer, quinic acid, nor other enzymes encoded in the qa gene cluster are necessary for the expression of activity. Therefore the combined genetic and biochemical data on the qa system continue to support the hypothesis that the qa-1 regulatory protein acts as a positive initiator of qa enzyme synthesis.
D000429:Alcohol Oxidoreductases / Q000096:biosynthesis; D000596:Amino Acids / Q000032:analysis; D005796:Genes; D005809:Genes, Regulator; D006836:Hydro-Lyases / Q000096:biosynthesis* / Q000276:immunology / Q000302:isolation & purification; D008970:Molecular Weight; D009154:Mutation; D009491:Neurospora / Q000201:enzymology*; D009492:Neurospora crassa / Q000201:enzymology* / Q000378:metabolism; D011801:Quinic Acid / Q000031:analogs & derivatives / Q000378:metabolism; D013045:Species Specificity; D013696:Temperature
D000596:Amino Acids; D011801:Quinic Acid; D000429:Alcohol Oxidoreductases; D006836:Hydro-Lyases