An NADP+-dependent glycerol dehydrogenase in Aspergillus nidulans is inducible by D-galacturonate

Sealy-Lewis, Heather M.; Fairhurst, Valerie

doi:10.1007/bf00317924

Cited by 18 publications

(18 citation statements)

References 16 publications

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“…ADHI, ADHII and ADHIII only exhibit activity in the gel stain when NAD' is present, and glycerol DHII only shows activity with NADP'. When NADP' was used as the electron acceptor with glycerol as the substrate, an activity band additional to glycerol DHII was seen which corresponded to the previously described glycerol DHI (Hondmann et al, 1991 ;Sealy-Lewis & Fairhurst, 1992). The latter enzyme is constitutively expressed and was previously shown to have activity with either glycerol or erythritol, but additionally it has some activity with 2,3-butanediol and 1,2-propanediol.…”

Section: Resultssupporting

confidence: 58%

Substrate specificity of nine NAD+-dependent alcohol dehydrogenases in Aspergillus nidulans

Sealy-Lewis

Fairhurst

1995

Microbiology

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In Aspergillus nidulans three alcohol dehydrogenases (ADHs) have been described. ADHI is induced by ethanol and is the physiological enzyme of ethanol utilization, ADHll has not been attributed a function but is repressed by ethanol. The ALCR regulatory protein acts positively to induce ADHI, and negatively in its control of ADHII. ADHlll is specifically induced by anaerobic stress. We have characterized the substrate specificity of these three enzymes by looking at their staining profile on polyacrylamide gels with a range of alcohols. In addition to these enzymes we have observed six other NAD+-dependent ADHs, two of which, propan-2-01 dehydrogenase and pentan-2-01 dehydrogenase, share similar control with ADHII. The inducibility of these enzymes with some alcohols has also been investigated. The profile of ADHs with NADP' as an electron acceptor is also reported.

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Section: Resultssupporting

confidence: 58%

Substrate specificity of nine NAD+-dependent alcohol dehydrogenases in Aspergillus nidulans

Sealy-Lewis

Fairhurst

1995

Microbiology

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“…Polyacrylamide Gel Electrophoresis and ADHI Activity-Gel electrophoresis and activity staining of ADHI were according to the method described by Sealy-Lewis and Fairhurst (20 The XbaI-XbaI fragment extracted from the pAN923-42BglII, carrying a mutant argB allele was cloned into the SmaI site of the bA2 plasmid, resulting in plasmid bA2 argB. The plasmids mutated in the AlcR-binding sites on the alcA promoter were generated by oligonucleotide-directed mutagenesis carried out by the method of Kunkel et al (22).…”

Section: Methodsmentioning

confidence: 99%

“…Independent experiments including mycelium growth and running gels were performed at least twice. Activity staining of ADHI was according to Sealy-Lewis and Fairhurst (20). The scanning diagram was normalized to a value 100, representing ADHI activity of the wild-type strain under these conditions.…”

Section: Alcr Binds In Vitro To Three Specific Targets In the Alcamentioning

confidence: 99%

The Zinc Binuclear Cluster Activator AlcR Is Able to Bind to Single Sites but Requires Multiple Repeated Sites for Synergistic Activation of the alcA Gene in Aspergillus nidulans

Panozzo

Capuano

Fillinger

et al. 1997

Journal of Biological Chemistry

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The alcA gene which is part of the recently identified ethanol regulon, is one of the most strongly inducible genes in Aspergillus nidulans. Its transcriptional activation is mediated by the AlcR transactivator which contains a DNA-binding domain belonging to the C6 zinc binuclear cluster family. AlcR differs from the other members of this family by several features, the most striking characteristic being its binding to both symmetric and asymmetric DNA sites with the same apparent affinity. However, AlcR is also able to bind to a single site with high affinity, suggesting that unlike the other C6 proteins, AlcR binds as a monomer.In this report, we show that AlcR targets, to be functional in vivo, have to be organized as inverted or direct repeats. In addition, we show a strong synergistic activation of alcA transcription in which the number and the position of the AlcR-binding sites are crucial. The fact that the AlcR unit for in vitro binding is a single site whereas the in vivo functional unit is a repeat opens the question of the mechanism of the strong alcA transactivation. These results show that AlcR displays both in vitro and in vivo a new range of binding specificity and provides a novel example in the C6 zinc cluster protein family.

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“…Polyacrylamide Gel Electrophoresis and ADHI Activity-Gel electrophoresis and activity staining of ADHI were carried out according to the method described by Sealy-Lewis and Fairhurst (27). Protein concentration was measured according to Bradford (28).…”

Section: Methodsmentioning

confidence: 99%

“…1, seven (27). The scanning diagram was normalized to a value of 10, representing ADHI activity of the wild type strain under ethanol-induced growth conditions.…”

Section: Identification Of Crea Binding Sites In the Alca Promoter-mentioning

confidence: 99%

The CreA Repressor Is the Sole DNA-binding Protein Responsible for Carbon Catabolite Repression of the alcA Gene in Aspergillus nidulans via Its Binding to a Couple of Specific Sites

Panozzo¹,

Cornillot

Felenbok

1998

Journal of Biological Chemistry

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Carbon catabolite repression is mediated in Aspergillus nidulans by the negative acting protein CreA. The CreA repressor plays a major role in the control of the expression of the alc regulon, encoding proteins required for the ethanol utilization pathway. It represses directly, at the transcriptional level, the specific transacting gene alcR, the two structural genes alcA and aldA, and other alc genes in all physiological growth conditions. Among the seven putative CreA sites identified in the alcA promoter region, we have determined the CreA functional targets in AlcR constitutive and derepressed genetic backgrounds. Two different divergent CreA sites, of which one overlaps a functional AlcR inverted repeat site, are largely responsible for alcA repression. Totally derepressed alcA expression is achieved when these two CreA sites are disrupted in addition to another single site, which overlaps the functional palindromic induction target. The fact that derepression is always associated with alcA overexpression is consistent with a competition model between AlcR and CreA for their cognate targets in the same region of the alcA promoter.Our results also indicate that the CreA repressor is necessary and sufficient for the total repression of the alcA gene.

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An NADP+-dependent glycerol dehydrogenase in Aspergillus nidulans is inducible by D-galacturonate

Cited by 18 publications

References 16 publications

Substrate specificity of nine NAD+-dependent alcohol dehydrogenases in Aspergillus nidulans

Substrate specificity of nine NAD+-dependent alcohol dehydrogenases in Aspergillus nidulans

The Zinc Binuclear Cluster Activator AlcR Is Able to Bind to Single Sites but Requires Multiple Repeated Sites for Synergistic Activation of the alcA Gene in Aspergillus nidulans

The CreA Repressor Is the Sole DNA-binding Protein Responsible for Carbon Catabolite Repression of the alcA Gene in Aspergillus nidulans via Its Binding to a Couple of Specific Sites

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