The Saccharomyces cerevisiae ADR1 gene is a positive regulator of transcription of genes encoding peroxisomal proteins.

Expression of the CTA1 gene of Saccharomyces cerevisiae, encoding catalase A, the peroxisomal catalase of this yeast, is sensitive to glucose repression. A DNA fragment cloned as a multicopy plasmid suppressing the glucose repression of CTA1 transcription was demonstrated to contain the ADR1 gene. Multiple copies of ADR1 increased catalase A formation not only on 10% glucose, but also on ethanol medium and in the presence of oleic acid, an inducer of peroxisome proliferation. Compared with wild-type cells, adr1 null mutants produced by disruption of the gene exhibit reduced CTA1 expression. This demonstrates that ADR1 is a true positive regulator of CTA1. Further experiments showed that it acts directly on CTA1. Alcohol dehydrogenase II, which is under ADR1 control, was excluded as a mediator of the effect on CTA1; deletion of bases -123 to -168 of CTA1 reduces expression and eliminates the response to the ADR1 multicopy plasmid without eliminating fatty acid induction; and gel retardation experiments demonstrated that ADR1 binds to a CTA1 upstream fragment (-156 to -184) with limited similarity to the ADR1 binding site of ADH2. Northern hybridization experiments further demonstrated that expression of two genes encoding enzymes of peroxisomal beta-oxidation (beta-ketothiolase, trifunctional enzyme) and of a gene involved in peroxisome assembly (PAS1) is also negatively affected by the adr1 null mutation. These findings demonstrate that the ADR1 protein has much broader regulatory functions than previously recognized.

A Saccharomyces cerevisiae upstream activating sequence mediates induction of peroxisome proliferation by fatty acids.

Peroxisome proliferation in Saccharomyces cerevisiae is induced by fatty acids via as yet unknown mechanisms. We have initiated a study of these mechanisms by identifying control sequences sufficient for fatty acid control of the CTA1 gene (encoding the peroxisomal catalase A). Promoter regions previously shown to be necessary for control were tested for their potential to mediate induction by oleic acid to a CYC1::lacZ fusion gene. A region previously demonstrated to control CTA1 via the ADR1 transcription activator (bp -156 to -184) does not mediate induction by oleic acid. In contrast, an adjacent sequence (-184 to -198) is sufficient for oleic acid induction, and a neighbouring element (-197 to -215) has marginal inducing activity. These two elements are characterized by a consensus sequence, 5'-CGGNNNTNA ('peroxisome box'), which is found in a number of S. cerevisiae peroxisomal protein-encoding genes. Mutation of either the CGG or the TNA block in the box has a dramatic down-regulating effect on the gene expression in oleic acid medium.

Sequence of the Saccharomyces cerevisiae CTA1 gene and amino acid sequence of catalase A derived from it.

The nucleotide sequence of a 2785-base-pair stretch of DNA containing the Saccharomyces cerevisiae catalase A (CTA1) gene has been determined. This gene contains an uninterrupted open reading frame encoding a protein of 515 amino acids (relative molecular mass 58,490). Catalase A, the peroxisomal catalase of S. cerevisiae was compared to the peroxisomal catalases from bovine liver and from Candida tropicalis and to the non-peroxisomal, presumably cytoplasmic, catalase T of S. cerevisiae. Whereas the peroxisomal catalases are almost colinear, three major insertions have to be introduced in the catalase T sequence to obtain an optimal fit with the other proteins. Catalase A is most closely related to the C. tropicalis enzyme. It is also more similar to the bovine liver catalase than to the second S. cerevisiae catalase. The differences between the two S. cerevisiae enzymes are most striking within four blocks of amino acids consisting of a total of 37 residues with high homology between the three peroxisomal, but low conservation between the S. cerevisiae catalases. The results obtained indicate that the peroxisomal catalases compared have very similar three-dimensional structures and might have similar targeting signals.