Alcohol oxidase expressed under nonmethylotrophic conditions is imported, assembled, and enzymatically active in peroxisomes of Hansenula polymorpha.

Distel, Ben; Ley, Inge van der; Veenhuis, Marten; Tabak, Henk F.

doi:10.1083/jcb.107.5.1669

Cited by 41 publications

(19 citation statements)

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“…This view is based on findings, obtained with overproducing transformants of H. polymorpha (14) and Saccharomyces cerevisiae (6), which indicated that peroxisomes have a considerably higher storage capacity than is normally utilized in wild-type cells (14). Therefore, our (3). Therefore, the finding that, besides AO, dihydroxyacetone synthase and thiolase protein are also present in the aggregates in C. boidinii suggests that in the early adaptation period, competition may exist for a common recognition and/or translocation machinery.…”

Section: Resultsmentioning

confidence: 96%

Development of multipurpose peroxisomes in Candida boidinii grown in oleic acid-methanol limited continuous cultures

et al. 1992

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We have studied the development and metabolic significance of peroxisomes in the yeast Candida boidinii following adaptation of the organism to cultivation conditions which require the simultaneous presence and activity of two independent peroxisome-mediated pathways for growth. After the addition of methanol to oleic acid-grown cells at late exponentional growth, a number of new small peroxisomes developed which, apart from the presence of 1-oxidation enzymes, were characterized by the presence of enzymes involved in methanol metabolism (alcohol oxidase and dihydroxyacetone synthase). The latter proteins, however, were absent in the larger organelles which were originally present in the oleic acid-grown cells prior to the addition of methanol and which contained only enzymes of the 13-oxidation pathway. Subsequent experiments on cells from continuous cultures grown on a mixture of oleic acid and methanol at steady-state conditions revealed that both the enzymes of the 13-oxidation pathway and those involved in methanol metabolism were found in one and the same compartment. Thus, under these conditions the cells contained peroxisomes which were concurrently involved in the metabolism of two different carbon sources simultaneously used for growth. Our results indicated that the heterogeneity in the peroxisomal population of a single cell, observed in the transient state following the addition of methanol, is only temporary and due to heterogeneity among these organelles with respect to their capacity to incorporate newly synthesized matrix proteins.In yeasts, microbodies (peroxisomes and glyoxysomes) play an essential role in the metabolism of different carbon and/or organic nitrogen sources (21,22). Initially, it was thought that the organelles were of minor metabolic significance and contained only one or more catalytic functions. The finding, however, that microbodies present in methylotrophic yeasts contain not only alcohol oxidase, the key enzyme of methanol metabolism, but also dihydroxyacetone synthase (4, 7), a key enzyme in carbon assimilation under these conditions (18), together with other biosynthetic enzymes (28) stresses the importance of these organelles and clearly places their metabolic significance beyond that of a compartment purely involved in catabolism.Yeast microbodies do not arise de novo but develop by growth and fission of existing organelles (22, 23); however, experiments performed with the methylotrophic yeast Hansenula polymorpha indicated that the capacity of individual organelles to incorporate newly synthesized matrix proteins may be related to their developmental stage (24).The recent finding that, in the methylotrophic yeast Candida boidinii, massive microbody proliferation may be induced by carbon sources other than methanol (9, 16) makes it possible to study the development and functioning of the microbodies during the growth of cells in media supplemented with more than one carbon source, each of which is metabolized via specific peroxisome-mediated metabolic pathways. The pre...

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Section: Resultsmentioning

confidence: 96%

Development of multipurpose peroxisomes in Candida boidinii grown in oleic acid-methanol limited continuous cultures

et al. 1992

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“…For instance, H. polymorpha pim mutants that are affected in matrix protein import contain enhanced numbers of small-sized peroxisomes relative to wild type organelles (33). On the other hand, overproduction of AO protein in glucose-grown cells resulted in enlarged organelles but not an increase in organelle numbers (34).…”

Section: Mpp1p Is a Member Of The Zn(ii) 2 Cys 6 Cluster Protein Famimentioning

confidence: 99%

Transcriptional Down-regulation of Peroxisome Numbers Affects Selective Peroxisome Degradation in Hansenula polymorpha

Leão-Helder

Krikken

Klei

et al. 2003

Journal of Biological Chemistry

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We have isolated and characterized a novel transcription factor of Hansenula polymorpha that is involved in the regulation of peroxisomal protein levels. This protein, designated Mpp1p, belongs to the family of Zn(II) 2 Cys 6 proteins. In cells deleted for the function of Mpp1p the levels of various proteins involved in peroxisome biogenesis (peroxins) and function (enzymes) are reduced compared with wild type or, in the case of the matrix protein dihydroxyacetone synthase, fully absent. Also, upon induction of mpp1 cells on methanol, the number of peroxisomes was strongly reduced relative to wild type cells and generally amounted to one organelle per cell. Remarkably, this single organelle was not susceptible to selective peroxisome degradation (pexophagy) and remained unaffected during exposure of methanol-induced cells to excess glucose conditions. We show that this mechanism is a general phenomenon in H. polymorpha in the case of cells that contain only a single peroxisome.Eukaryotic cells are thought to have evolved ϳ1.5 billion years ago. The development of cell organelles allowed primitive eukaryotes to compartmentalize specific cellular functions. Concomitantly, genetic mechanisms that control the biogenesis and function of these compartments had to be developed. Obviously, the separate classes of organelles are characterized by their specific function, as in energy metabolism (mitochondrion), degradation processes (vacuole, lysosome), or protein transport (Golgi system, endoplasmatic reticulum).Among the organelles, peroxisomes are remarkable because of their highly versatile functions most of which are related to specific metabolic pathways in the organism in which they occur. This functional flexibility is not reflected in their morphology. The organelles are invariably very simple of construction and consist of a proteinaceous matrix, surrounded by a single membrane. Nevertheless, their function varies from the oxidation of very long chain fatty acids in man, germination of oil-bearing seed and photorespiration in green plants, to the metabolism of unusual carbon and/or nitrogen sources in fungi (1). In the methylotrophic yeast Hansenula polymorpha peroxisomes are essential to support growth of cells on media containing methanol as the sole source of carbon and energy. Under these conditions many organelles that contain the key enzymes involved in methanol metabolism, alcohol oxidase (AO), 1 dihydroxyacetone synthase (DHAS), and catalase, develop in the cells. Conversely, when methanol-grown wild type (WT) cells are shifted to conditions in which the organelles are redundant for growth (e.g. glucose), they are rapidly and sequentially degraded by a process designated pexophagy (reviewed in Ref. 2). Morphological data suggest that in each cell generally a single (or few) small peroxisome(s) escape(s) the degradation process. The resistance of these organelles to degradation is thought to be of physiological advantage in that it allows the cells to quickly adapt to new environments that require new per...

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“…For instance, in the yeast Succhuromyces cerevisiue few peroxisomes are present during growth on glucose and they proliferate when cells are shifted to oleic acid as the sole carbon source [7,8]. This proliferation of peroxisomes cannot be triggered by an increase in the volume of the original organelle due to the over-production of one peroxisomal matrix protein, but seems to be regulated otherwise, as has been shown by Distel et al [9] and Godecke et al [lo]. In order to gain more insight into peroxisome biogenesis, we have initiated the study of transcriptional regulation of genes coding for peroxisomal proteins in S. cerevisiae.…”

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confidence: 92%

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

Einerhand

Voorn-Brouwer

Erdmann

et al. 1991

European Journal of Biochemistry

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Transferring Succhuromyces cerevisiae cells from glucose-to oleate-containing growth media results in a significant increase in the number and volume of peroxisomes. To investigate this proliferation process we studied the transcriptional regulation of the gene coding for peroxisomal3-oxoacyl-CoA thiolase (EC 2.3.1.16) in response to the switch in carbon source. Expression was proved to be repressed during growth on glucose, derepressed during growth on glycerol, and induced during growth on oleate as the sole carbon source. By deletion and mutational analysis of sequences upstream of this gene, we have identified a region which is involved in the regulation of transcription. It is contained within a 52-base-pair sequence, UASTs2 (upstream activation sequence thiolase 52), located between 203 and 151 nucleotides upstream of the translational initiation codon. This sequence proved to be required for repression, derepression and induction of transcription, and was able to activate transcription from the truncated version of the heterologous iso-1-cytochrome-c (CYCI) promoter in a similar way as in the wild-type promoter context.Sequence comparison revealed that the UASTs2 contained a sequence motif ('P-oxidation box') that is very similar to sequences located in the 5'-upstream regions of the genes coding for two other /-oxidation enzymes of S. cerevisiue : the peroxisomal acyl-CoA oxidase and the peroxisomal trifunctional P-oxidation enzyme of S. cerevisiue. Mutational analysis of the 'P-oxidation box' indicates that this sequence motif acts as a UAS in vivo.Sequence comparison also revealed that just upstream of the 'P-oxidation box', between positions -21 3 and -201, a potential binding site occurred for the yeast multifunctional autonomously replicating sequence binding factor ABFl . Gel-retardation-competition experiments indicate that ABFl binds specifically to this sequence.The peroxisome is a ubiquitous organelle present in almost all eukaryotes (see [l] for a recent review). It is defined as a subcellular organelle containing catalase and at least one hydrogen-peroxide-producing oxidase [2]. Depending on the organism, peroxisomes function in the catabolism of a wide variety of substrates such as fatty acids, D-amino acids, methanol, L-a-hydroxy acids, uric acid and polyamines. They also play a crucial role in the biosynthesis of ether-linked glycerolipids such as plasmalogens, in the metabolism of cholesterol and phytanic acid, and in the synthesis of bile acids [3, 41. Many lines of biochemical and morphological evidence indicate that peroxisomes are formed by division of pre-existing peroxisomes after post-translational import of newly synthesized proteins (see [S, 61 for reviews). Peroxisomal proteins, including membrane proteins, are synthesized on free polyribosomes in the cytosol, mostly at their mature sizes.

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Alcohol oxidase expressed under nonmethylotrophic conditions is imported, assembled, and enzymatically active in peroxisomes of Hansenula polymorpha.

Cited by 41 publications

References 36 publications

Development of multipurpose peroxisomes in Candida boidinii grown in oleic acid-methanol limited continuous cultures

Development of multipurpose peroxisomes in Candida boidinii grown in oleic acid-methanol limited continuous cultures

Transcriptional Down-regulation of Peroxisome Numbers Affects Selective Peroxisome Degradation in Hansenula polymorpha

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

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