Summary: Aspergillus niger mutants relieved of carbon repression were isolated from an areA parental strain by selection of colonies that exhibited improved growth on a combination of 4-aminobutanoic acid (GABA) and D-glucose. In addition to derepression of the utilization of GABA as a nitrogen source in the presence of D-glucose, three of the four mutants also showed derepression of L-alanine and L-proline utilization. Transformation of the mutants with the A. niger creA gene, encoding the repressor protein CREA, re-established the areA phenotype on GABA/D-glucose, identifying the mutations as creAd. The creA gene mapped on chromosome IV by linkage analysis and contour-clamped homogeneous electric field hybridization. The creA mutants obtained were used to study the involvement of CREA in repression by D-glucose of arabinases and L-arabinose catabolism in A. niger. In wild-type A. niger, α-L-arabinofuranosidase A, α-L-arabinofuranosidase B, endo-arabinase, L-arabinose reductase and L-arabitol dehydrogenase were induced on L-arabinose, but addition of D-glucose prevented this induction. Repression was relieved to varying degrees in the creA mutants, showing that biosynthesis of arabinases and L-arabinose catabolic enzymes is under control of CREA.
ArstH. N.Jr,
BaileyC. R.1977; The regulation of carbon metabolism in Aspergillus nidulans
. In Genetics and Physiology of Aspergillus nidulans pp. 131–146 Edited by
SmithJ. E.,
PatemanJ. A.
London: Academic Press;
BosC. J.,
DebetsA. J. M.,
SwartK.,
HuybersA.,
KobusG.,
SlakhorstS. M.1988; Genetic analysis and the construction of master strains for assignment of genes to six linkage groups in Aspergillus niger
. Curr Genet 14:437–443
CuberoB.,
ScazzocchioC.1994; Two different, adjacent and divergent zinc finger binding sites are necessary for CREA-mediated carbon catabolite repression in the proline gene cluster of Aspergillus nidulans
. EMBO J 13:407–415
DowzerC. E. A.,
KellyJ. M.1991; Analysis of the creA gene, a regulator of carbon catabolite repression in Aspergillus nidulans
. Mol Cell Biol 11:5701–5709
FillingerS.,
FelenbokB.1996; A newly identified gene cluster in Aspergillus nidulans comprises five novel genes localized in the alc region that are controlled both by the specific transactivator AlcR and the general carbon-catabolite repressor CreA. Mol Microbiol 20:475–488
FlipphiM. J. A.,
van HeuvelM.,
Van der VeenP.,
VisserJ.,
de GraaffL. H.1993a; Cloning and characterization of the abfB gene coding for the major α-l-arabinofuranosidase (ABF B) of Aspergillus niger
. Curr Genet 24:525–532
FlipphiM. J. A.,
PannemanH.,
Van der VeenP.,
VisserJ.,
de GraaffL. H.1993b; Molecular cloning, expression and structure of the endo-l,5-α-l-arabinase gene of Aspergillus niger
. Appl Microbiol Biotechnol 40:318–326
FlipphiM. J. A.,
VisserJ.,
Van der VeenP.,
de GraaffL. H.1993c; Cloning of the Aspergillus niger gene encoding α-l-arabinofuranosidase A. Appl Microbiol Biotechnol 39:335–340
FlipphiM. J. A.,
VisserJ.,
Van der VeenP.,
de GraaffL. H.1994; Arabinase gene expression in Aspergillus niger: indications for coordinated regulation. Microbiology 140:2673–2682
GoosenT.,
BloemheuvelG.,
GyslerC.,
de BieD. A.,
Van den BroekH. W. J.,
SwartK.1987; Transformation of Aspergillus niger using the homologous orotidine-5ʹ-phosphate-decarboxy-lase gene. Curr Genet 11:499–503
JohnstonM.,
FlickJ. S.,
PextonT.1994; Multiple mechanisms provide rapid and stringent glucose repression of gal gene expression in Saccharomyces cerevisiae
. Mol Cell Biol 14:3834–3841
KulmburgP.,
MathieuM.,
Dowzer, G, KellyJ.,
FelenbokB.1993; Specific binding sites in the alcR and alcA promoters of the ethanol regulon for the CREA repressor mediating carbon catabolite repression in Aspergillus nidulans
. Mol Microbiol 7:847–857
Kusters-van SomerenM. A.,
HarmsenJ. A. M.,
KesterH. C. M.,
VisserJ.1991; Structure of the Aspergillus niger pelA. gene and its expression in Aspergillus niger and Aspergillus nidulans
. Curr Genet 20:293–299
NehlinJ. O.,
CarlbergM.,
RonneH.1991; Control of yeast GAL genes by MIG1 repressor: a transcriptional cascade in the glucose response. EMBO J 10:3373–3377
SchaapP. J.,
MüllerY.,
BaarsJ. J. P.,
Op den CampH. J. M.,
SonnenbergA. S. M.,
Van GriensvenL. J. L.,
D. & VisserJ.1996; Nucleotide sequence and expression of the gene encoding NADP+-dependent glutamate dehydrogenase (gdhk) from Agaricus bisporus
. Mol Gen Genet 250:339–347
ShroffR. A.,
LockingtonR. A.,
KellyJ. M.1996; Analysis of mutations in the creA gene involved in carbon catabolite repression in Aspergillus nidulans
. Can J Microbiol 42:950–959
Van der VeenP.,
FlipphiM. J. A.,
VoragenA. G. J.,
VisserJ.1991; Induction, purification and characterisation of arabinases produced by Aspergillus niger
. Arch Microbiol 157:23–28
Van der VeenP.,
FlipphiM. J. A.,
VoragenA. G. J.,
VisserJ.1993; Induction of extracellular arabinases on monomeric substrates in Aspergillus niger
. Arch Microbiol 159:66–71
Van der VeenP.,
ArstH. N.,
Jr, FlipphiM. J. A.,
VisserJ.1994; Extracellular arabinases in Aspergillus nidulans: the effect of different cre mutations on enzyme levels. Arch Microbiol 162:433–440
VerdoesJ. C.,
CalilM. R.,
PuntP. J.,
DebetsF.,
SwartK.,
StouthamerA. H.,
Van den HondelC. A. J. J.1994; The complete karyotype of Aspergillus niger: the use of introduced electrophoretic mobility variation of chromosomes tor gene assignment studies. Mol Gen Genet 244:75–80
WitteveenC. F. B.,
BusinkR.,
Van de VondervoortP. J. I.,
DijkemaC.,
SwartK.,
VisserJ.1989; l-Arabinose and d-xylose catabolism in Aspergillus niger
. J Gen Microbiol 135:2163–2171
WitteveenC. F. B.,
WeberF.,
BusinkR.,
VisserJ.1994; Isolation and characterisation of two xylitol dehydrogenases from Aspergillus niger
. Microbiology 140:1679–1685