Abstract
Complementation by transformation of an Aspergillus niger mutant lacking xylanolytic activity led to the isolation of the xlnR gene. The xlnR gene encodes a polypeptide of 875 amino acids capable of forming a zinc binuclear cluster domain with similarity to the zinc clusters of the GAL4 superfamily of transcription factors. The XlnR-binding site 5'-GGCTAAA-3' was deduced after electrophoretic mobility shift assays, DNase I footprinting and comparison of various xylanolytic promoters. The importance of the second G within the presumed XlnR binding site 5'-GGCTAAA-3' was confirmed in vitro and in vivo. The 5'-GGCTAAA-3' consensus sequence is found within several xylanolytic promoters of various Aspergillus species and Penicillium chrysogenum. Therefore, this sequence may be an important and conserved cis-acting element in induction of xylanolytic genes in filamentous fungi. Our results indicate that XlnR is a transcriptional activator of the xylanolytic system in A. niger.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Aspergillus niger / enzymology
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Aspergillus niger / genetics*
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Aspergillus niger / metabolism*
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Base Sequence
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Binding, Competitive
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Blotting, Northern
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Blotting, Southern
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Blotting, Western
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DNA Footprinting
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DNA, Fungal / chemistry
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Deoxyribonuclease I
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Electrophoresis, Polyacrylamide Gel
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Fungal Proteins*
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Gene Expression Regulation, Enzymologic / physiology
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Gene Expression Regulation, Fungal / physiology*
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Molecular Sequence Data
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Mutation
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Promoter Regions, Genetic / genetics
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Restriction Mapping
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Sequence Analysis, DNA
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Trans-Activators / genetics
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Transcription, Genetic
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Transformation, Genetic
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Xylan Endo-1,3-beta-Xylosidase
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Xylans / metabolism*
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Xylosidases / chemistry
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Xylosidases / genetics*
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Xylosidases / metabolism
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Zinc Fingers / genetics
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Zinc Fingers / physiology
Substances
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DNA, Fungal
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Fungal Proteins
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Trans-Activators
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XlnR protein, Aspergillus
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Xylans
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Deoxyribonuclease I
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Xylosidases
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Xylan Endo-1,3-beta-Xylosidase