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

Abstract

The nucleotide sequence of a 2785-base-pair stretch of DNA containing the Saccharomyces cerevisiae catalase A (CTAl) gene has been determined. This gene contains an uninterrupted open reading frame encoding a protein of 515 amino acids (relative molecular mass 58490). Catalase A, the peroxisomal catalase of S. cerevisiae was compared to the peroxisomal catalases from bovine liver and from Cundidu tropicalis and to the non-peroxisomal, presumably cytoplasmic, catalase T of S. cerevisiue. 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. cerevisiue catalase. The differences between the two S. cerevisiue 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. cerevisiue catalases. The results obtained indicate that the peroxisomal catalases compared have very similar three-dimensional structures and might have similar targetting signals.Catalases are components of a system of enzymes functioning in the detoxification of oxygen metabolites formed in a variety of biological processes [I -31. Accordingly they are present in most prokaryotic and eukaryotic organisms. Catalase from mammalian liver has been the subject of many biochemical studies [4]. Bovine liver catalase has been studied extensively by X-ray crystallography [5 -71 and its three-dimensional structure has been compared to that of the catalase from Penicillium vitale [8]. Regulation of catalase synthesis has also been studied systematically in a number of organisms [9-151. Although catalases have been the subject of many investigations, important aspects of their functions, of the relationship between the structure of the proteins and their catalytic activity, of the control of biosynthesis and of the intracellular transport of this enzyme are only poorly understood. The yeast Saccharomyces cerevisiue is an organism particularly well-suited for studying such aspects by genetic and biochemical methods. DNA sequencing A 4.5-kb HindIII fragment, containing the entire catalase A gene and its flanking regions, was cloned in plasmid pUCl9. HindIII-BumHI, HindIII-KpnI and HindIII-HpaI subfragments derived from the HindIII fragment were also cloned in pUC19. AluI, HueIII, Suu3A and TuqI fragments were cloned in plasmid pUCl8. Double-stranded plasmids were isolated by the method of Birnboim and Doly [25]. The inserts were sequenced by the dideoxy-chain-termination method [26] using labelled universal or specific primers basically as described by Chen and Seeburg [27] and by Korneluk et al. [28]. The right and the left ends of the 2.8-kb EcoRI fragment were sequenced using the dideoxy-chain-termination method after cloning the fragment in both orientations into M13 mp18 [24]....

Regulation of transcription of the gene coding for peroxisomal 3‐oxoacyl‐CoA thiolase of Saccharomcyes cerevisiae

“…SA, B) was taken as 100% pecially highly conserved. Furthermore, in the 5'-upstream region of the gene coding for the oleate-inducible catalase A of S. cerevisiae the sequence between -21 1 and -203 shows a significant (78%) similarity to this conserved part of the 'Poxidation box' [46]. These 'P-oxidation boxes' or part of them could therefore be potential binding sites for trans-acting factors responsible for the oleate-specific induction effect.…”

Characterization of a transcriptional control element involved in proliferation of peroxisomes in yeast in response to oleate

“…7; Wang et al, 1992;Hiltunen et al, 1992;Cohen et al, 1988;., 1991). In (Dmochowska et al, 1990); FOX2 gene, encoding the yeast multifunctional enzyme (Hiltunen et al, 1992); FOX3 gene, encoding the yeast thiolase gene (Einerhand et al, 1991); CTAl gene, encoding the peroxisomal catalase A (Cohen et al, 1988); PASl gene, encoding a protein involved in peroxisomal assembly . The sequences are positioned so that maximal sequence similarity is obtained.…”

“…Sequence comparison revealed that the 5' flanking regions of several other genes, encoding proteins involved in peroxisomal function or assembly (POX1 = FOXl, FOX2, CTAl and PAS1 genes), also contain similar palindromic sequence motifs in their promoter regions ( Fig. 7; Wang et al, 1992;Hiltunen et al, 1992;Cohen et al, 1988;., 1991). In (Dmochowska et al, 1990); FOX2 gene, encoding the yeast multifunctional enzyme (Hiltunen et al, 1992); FOX3 gene, encoding the yeast thiolase gene (Einerhand et al, 1991); CTAl gene, encoding the peroxisomal catalase A (Cohen et al, 1988); PASl gene, encoding a protein involved in peroxisomal assembly .…”

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

“…The yeast Saccharomyces cerevisiae is an ideal system for studying such mechanisms because it allows the use of an efficient combination of methods of cell biology and genetics. The S. cerevisiae CTAI gene, which encodes catalase A, a peroxisomal protein (31), has been cloned and sequenced (13,14). Catalase A has been demonstrated to be induced by fatty acids together with peroxisomal structures and a number of other peroxisomal proteins (31,39).…”