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3. Regulation of acid phosphatases in anAspergillus niger pacC disruption strain

Author

P.J.I. van de Vondervoort, Andrew MacCabe, J. P. T. W. van den Hombergh, and Jaap Visser

Subjects
Molecular Genetics of Industrial Micro-organisms, Phosphate repression, Acid Phosphatase, Molecular Sequence Data, Phosphatase, Gene disruption, Fungal Proteins, Moleculaire genetica van industriële micro-organismen, Gene Expression Regulation, Fungal, Genes, Regulator, Genetics, Amino Acid Sequence, RNA, Messenger, DNA, Fungal, Molecular Biology, VLAG, Regulator gene, Regulation of gene expression, Fungal protein, biology, Aspergillus niger, Fungal genetics, Acid phosphatase, Chromosome Mapping, RNA, Fungal, Hydrogen-Ion Concentration, biology.organism_classification, Molecular biology, Biochemistry, biology.protein, Enzyme localization, Alkaline phosphatase, Transformation, Bacterial, Transcription Factors
Abstract

An Aspergillus niger strain has been constructed in which the pH-dependent regulatory gene, pacC, was disrupted. The pacC gene of A. niger, like that of A. nidulans, is involved in the regulation of acid phosphatase expression. Disruptants were identified by a reduction in acid phosphatase staining of colonies. Southern analysis demonstrated integration of the disruption plasmid at the pacC locus and Northern analysis showed that the disruption strain produced a truncated pacC mRNA of 2.2 kb (as compared to 2.8 kb in the wild type). The strain carrying the pacC disruption was used to assign the pacC gene to linkage group IV; this was confirmed by CHEF electrophoresis and Southern analysis. This strain further allowed us to determine which extracellular enzyme and transport systems are under the control of pacC in A. niger. Expression of the A. niger pacC wild-type gene and the truncated pacC gene showed that, in contrast to the auto-regulated wild-type expression, which was elevated only at alkaline pH, the truncated pacC gene was deregulated, as high-level expression occurred regardless of the pH of the culture medium. Analysis of the phosphatase spectrum by isoelectric focussing and enzyme activity staining both in the wild-type and the pacC disruptant showed that at least three acid phosphatases are regulated by the pacC. For the single alkaline phosphatase no pH regulation was observed.

Published
1996

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