Regulation of acid phosphatases in anAspergillus niger pacC disruption strain
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AnAspergillus niger strain has been constructed in which the pH-dependent regulatory gene,pacC, was disrupted. ThepacC gene ofA. niger, like that ofA. 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 thepacC locus and Northern analysis showed that the disruption strain produced a truncatedpacC mRNA of 2.2 kb (as compared to 2.8 kb in the wild type). The strain carrying thepacC disruption was used to assign thepacC 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 ofpacC inA. niger. Expression of theA. niger pacC wild-type gene and the truncatedpacC gene showed that, in contrast to the auto-regulated wild-type expression, which was elevated only at alkaline pH, the truncatedpacC 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 thepacC disruptant showed that at least three acid phosphatases are regulated by thepacC. For the single alkaline phosphatase no pH regulation was observed.
Key wordsPhosphate repression Gene disruption Enzyme localization
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