Abstract
In Neurospora crassa the qa-2 gene, which encodes catabolic dehydroquinase, is under positive control exerted by the inducer quinic acid and an activator protein encoded in the closely linked qa-1 gene. In order to determine if this regulatory mechanism is maintained when the qa-2 gene is cloned on a recombinant plasmid and expressed in Escherichia coli, molecular cloning experiments have been performed using DNA isolated from a qa-1+ (inducible), a qa-1C (constitutive) and two qa-1 (non-inducible) strains of N. crassa. The results demonstrate that the level of expression of the qa-2 gene in E. coli is completely independent of the mutational state of the qa-1 gene. Moreover, the level of expression of the cloned qa-2 gene was unaffected by either an intracellularly produced inducer of catabolic dehydroquinase or by the general procaryotic positive effector, the CAP factor. The weight of evidence thus supports the conclusion that transcription of the N. crassa qa-2 gene in E. coli does not require the qa-1 activator protein and thus is not controlled by the same mecahnism which functions in N. crassa.
MeSH Terms
Centrifugation, Density Gradient
DNA, Recombinant
Electrophoresis
Escherichia coli/genetics
Genes
Hydro-Lyases/genetics
Neurospora/genetics
Neurospora crassa/genetics
Plasmids
Quinic Acid/analogs & derivatives
Transcription, Genetic
Chemicals
DNA, Recombinant
Quinic Acid
Hydro-Lyases
3-dehydroquinate dehydratase
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Hautala J A
Schweizer M
Giles N H
Kushner S R
References (18)
18 references, click to expand
-
Genetic mapping of xthA, the structural gene for exonuclease III in Escherichia coli K-12.
J Bacteriol. 1976 Jun;126(3):1082-8
PMID: 780339
-
Purification and characterization of catabolic dehydroquinase, an enzyme in the inducible quinic acid catabolic pathway of Neurospora crassa.
J Biol Chem. 1975 Aug 10;250(15):6008-14
PMID: 125280
-
Genetical and biochemical evidence for further interrelationships between the polyaromatic synthetic and the quinate-shikimate catabolic pathways in Neurospora crassa.
Genetics. 1972 Jul;71(3):337-48
PMID: 4261015
-
Functional genetic expression of eukaryotic DNA in Escherichia coli.
Proc Natl Acad Sci U S A. 1976 May;73(5):1471-5
PMID: 775490
-
The inducible quinate-shikimate catabolic pathway in Neurospora crassa: genetic organization.
J Gen Microbiol. 1974 Apr;81(2):337-55
PMID: 4275708
-
Expression in Escherichia coli K-12 of the structural gene for catabolic dehydroquinase of Neurospora crassa.
Proc Natl Acad Sci U S A. 1977 Aug;74(8):3508-12
PMID: 143663
-
Recombinant levels of Escherichia coli K-12 mutants deficient in various replication, recombination, or repair genes.
J Bacteriol. 1978 Jun;134(3):958-66
PMID: 350859
-
Mutants in the arom gene cluster of Neurospora crassa specific for biosynthetic dehydroquinase.
Genetics. 1969 Apr;61(4):789-800
PMID: 5364964
-
The expression of cloned eukaryotic DNA in prokaryotes.
Brookhaven Symp Biol. 1977 May 12-20;(29):277-96
PMID: 383204
-
Molecular cloning and expression in E. coli of a yeast gene coding for beta-galactosidase.
Cell. 1978 Sep;15(1):123-30
PMID: 100226
-
Recalibrated linkage map of Escherichia coli K-12.
Bacteriol Rev. 1976 Mar;40(1):116-67
PMID: 773363
-
Genetic engineering in vivo using translocatable drug-resistance elements. New methods in bacterial genetics.
J Mol Biol. 1977 Oct 15;116(1):125-59
PMID: 338917
-
Transcription and translation in E. coli of hybrid plasmids containing the catabolic dehydroquinase gene from Neurospora crassa.
Gene. 1978 Nov;4(3):241-59
PMID: 154430
-
Genetic evidence on the organization and action of the qa-1 gene product: a protein regulating the induction of three enzymes in quinate catabolism in Neurospora crassa.
Proc Natl Acad Sci U S A. 1975 Feb;72(2):553-7
PMID: 123642
-
Functional expression of cloned yeast DNA in Escherichia coli.
Proc Natl Acad Sci U S A. 1977 Feb;74(2):487-91
PMID: 322128
-
Constitutive mutants in a regulatory gene exerting positive control of quinic acid catabolism in Neurospora crassa.
Proc Natl Acad Sci U S A. 1971 Jul;68(7):1555-9
PMID: 5283945
-
Expression of the structural gene for catabolic dehydroquinase of Neurospora crassa in Escherichia coli K12.
Brookhaven Symp Biol. 1977 May 12-20;(29):297-308
PMID: 157795
-
Regulation: positive control.
Annu Rev Genet. 1974;8:219-42
PMID: 4374117