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1.
Biochim Biophys Acta ; 1763(12): 1647-54, 2006 Dec.
Article in English | MEDLINE | ID: mdl-17030445

ABSTRACT

Eukaryotic cells contain functionally distinct, membrane enclosed compartments called organelles. Here we like to address two questions concerning this architectural lay out. How did this membrane complexity arise during evolution and how is this collection of organelles maintained in multiplying cells to ensure that new cells retain a complete set of them. We will try to address these questions with peroxisomes as a focal point of interest.


Subject(s)
Peroxisomes/physiology , Phylogeny , Animals , Biological Evolution , Endoplasmic Reticulum/physiology , Humans
2.
J Cell Biol ; 155(6): 979-90, 2001 Dec 10.
Article in English | MEDLINE | ID: mdl-11733545

ABSTRACT

In vivo time-lapse microscopy reveals that the number of peroxisomes in Saccharomyces cerevisiae cells is fairly constant and that a subset of the organelles are targeted and segregated to the bud in a highly ordered, vectorial process. The dynamin-like protein Vps1p controls the number of peroxisomes, since in a vps1Delta mutant only one or two giant peroxisomes remain. Analogous to the function of other dynamin-related proteins, Vps1p may be involved in a membrane fission event that is required for the regulation of peroxisome abundance. We found that efficient segregation of peroxisomes from mother to bud is dependent on the actin cytoskeleton, and active movement of peroxisomes along actin filaments is driven by the class V myosin motor protein, Myo2p: (a) peroxisomal dynamics always paralleled the polarity of the actin cytoskeleton, (b) double labeling of peroxisomes and actin cables revealed a close association between both, (c) depolymerization of the actin cytoskeleton abolished all peroxisomal movements, and (d) in cells containing thermosensitive alleles of MYO2, all peroxisome movement immediately stopped at the nonpermissive temperature. In addition, time-lapse videos showing peroxisome movement in wild-type and vps1Delta cells suggest the existence of various levels of control involved in the partitioning of peroxisomes.


Subject(s)
Actins/metabolism , Carrier Proteins/metabolism , Cytoskeletal Proteins , GTP-Binding Proteins , Myosin Heavy Chains/metabolism , Myosin Type V/metabolism , Peroxisomes/metabolism , Saccharomyces cerevisiae Proteins/metabolism , Saccharomyces cerevisiae/metabolism , Actin Cytoskeleton/metabolism , Bacterial Proteins/genetics , Base Sequence , Carrier Proteins/genetics , Gene Deletion , Green Fluorescent Proteins , Indicators and Reagents/metabolism , Luminescent Proteins/genetics , Membrane Proteins/genetics , Microtubule-Associated Proteins/genetics , Microtubules , Molecular Sequence Data , Mutagenesis/physiology , Oligonucleotide Probes , Peroxisome-Targeting Signal 1 Receptor , Polymers/metabolism , Receptors, Cytoplasmic and Nuclear/genetics , Receptors, Cytoplasmic and Nuclear/metabolism , Saccharomyces cerevisiae/genetics , Vesicular Transport Proteins
3.
FEBS Lett ; 506(1): 73-8, 2001 Sep 28.
Article in English | MEDLINE | ID: mdl-11591374

ABSTRACT

We have studied the mechanisms that regulate the remodeling of the glycolytic, mitochondrial and structural network of muscles of creatine kinase M (M-CK)/sarcomeric mitochondrial creatine kinase (ScCKmit) knockout mice by comparison of wild-type and mutant mRNA profiles on cDNA arrays. The magnitudes of changes in mRNA levels were most prominent in M-CK/ScCKmit (CK(-/-)) double mutants but did never exceed those of previously observed changes in protein level for any protein examined. In gastrocnemius of CK(-/-) mice we measured a 2.5-fold increase in mRNA level for mitochondrial encoded cytochrome c oxidase (COX)-III which corresponds to the increase in protein content. The level of the nuclear encoded mRNAs for COX-IV, H(+)-ATP synthase-C, adenine nucleotide translocator-1 and insulin-regulatable glucose transporter-4 showed a 1.5-fold increase, also in agreement with protein data. In contrast, no concomitant up-regulation in mRNA and protein content was detected for the mitochondrial inorganic phosphate-carrier, voltage-dependent anion channel and certain glycolytic enzymes. Our results reveal that regulation of transcript level plays an important role, but it is not the only principle involved in the remodeling of mitochondrial and cytosolic design of CK(-/-) muscles.


Subject(s)
Adaptation, Physiological/genetics , Creatine Kinase/genetics , Isoenzymes/genetics , Muscle, Skeletal/metabolism , RNA, Messenger/genetics , Animals , Creatine Kinase, Mitochondrial Form , Mice , Mice, Inbred C57BL , Mice, Knockout , Muscle, Skeletal/physiology , Phenotype
4.
J Cell Sci ; 114(Pt 11): 2199-204, 2001 Jun.
Article in English | MEDLINE | ID: mdl-11493655

ABSTRACT

The classic model for peroxisome biogenesis states that new peroxisomes arise by the fission of pre-existing ones and that peroxisomal matrix and membrane proteins are recruited directly from the cytosol. Recent studies challenge this model and suggest that some peroxisomal membrane proteins might traffic via the endoplasmic reticulum to peroxisomes. We have studied the trafficking in human fibroblasts of three peroxisomal membrane proteins, Pex2p, Pex3p and Pex16p, all of which have been suggested to transit the endoplasmic reticulum before arriving in peroxisomes. Here, we show that targeting of these peroxisomal membrane proteins is not affected by inhibitors of COPI and COPII that block vesicle transport in the early secretory pathway. Moreover, we have obtained no evidence for the presence of these peroxisomal membrane proteins in compartments other than peroxisomes and demonstrate that COPI and COPII inhibitors do not affect peroxisome morphology or integrity. Together, these data fail to provide any evidence for a role of the endoplasmic reticulum in peroxisome biogenesis.


Subject(s)
ATP-Binding Cassette Transporters , COP-Coated Vesicles/metabolism , Coat Protein Complex I/antagonists & inhibitors , Fungal Proteins , Membrane Proteins/metabolism , Peroxisomes/metabolism , Brefeldin A/pharmacology , COP-Coated Vesicles/drug effects , Cells, Cultured , Coat Protein Complex I/metabolism , Endoplasmic Reticulum/drug effects , Endoplasmic Reticulum/metabolism , Fibroblasts , Humans , Intracellular Membranes/drug effects , Intracellular Membranes/metabolism , Lipoproteins/biosynthesis , Lipoproteins/genetics , Lipoproteins/metabolism , Membrane Proteins/biosynthesis , Membrane Proteins/genetics , Microscopy, Fluorescence , Peroxins , Peroxisomal Biogenesis Factor 2 , Peroxisomes/drug effects , Protein Transport/drug effects
5.
Mol Cell Biol ; 21(13): 4321-9, 2001 Jul.
Article in English | MEDLINE | ID: mdl-11390660

ABSTRACT

We have characterized the role of YPR128cp, the orthologue of human PMP34, in fatty acid metabolism and peroxisomal proliferation in Saccharomyces cerevisiae. YPR128cp belongs to the mitochondrial carrier family (MCF) of solute transporters and is localized in the peroxisomal membrane. Disruption of the YPR128c gene results in impaired growth of the yeast with the medium-chain fatty acid (MCFA) laurate as a single carbon source, whereas normal growth was observed with the long-chain fatty acid (LCFA) oleate. MCFA but not LCFA beta-oxidation activity was markedly reduced in intact ypr128cDelta mutant cells compared to intact wild-type cells, but comparable activities were found in the corresponding lysates. These results imply that a transport step specific for MCFA beta-oxidation is impaired in ypr128cDelta cells. Since MCFA beta-oxidation in peroxisomes requires both ATP and CoASH for activation of the MCFAs into their corresponding coenzyme A esters, we studied whether YPR128cp is an ATP carrier. For this purpose we have used firefly luciferase targeted to peroxisomes to measure ATP consumption inside peroxisomes. We show that peroxisomal luciferase activity was strongly reduced in intact ypr128cDelta mutant cells compared to wild-type cells but comparable in lysates of both cell strains. We conclude that YPR128cp most likely mediates the transport of ATP across the peroxisomal membrane.


Subject(s)
Carrier Proteins/metabolism , Fatty Acids/metabolism , Fungal Proteins/metabolism , Nucleotide Transport Proteins , Peroxisomes/metabolism , Saccharomyces cerevisiae Proteins , Saccharomyces cerevisiae/physiology , Adenosine Triphosphate/metabolism , Carrier Proteins/chemistry , Carrier Proteins/genetics , Cell Fractionation , Fungal Proteins/chemistry , Fungal Proteins/genetics , Genes, Reporter/genetics , Glucose/metabolism , Humans , Immunoblotting , Lauric Acids/metabolism , Luciferases/genetics , Luciferases/metabolism , Membrane Proteins/chemistry , Membrane Proteins/genetics , Membrane Proteins/metabolism , Mitochondria/chemistry , Mitochondria/metabolism , Oleic Acid/metabolism , Oxidation-Reduction , Peroxisomes/chemistry , Peroxisomes/ultrastructure , Recombinant Fusion Proteins/genetics , Recombinant Fusion Proteins/metabolism , Saccharomyces cerevisiae/genetics , Saccharomyces cerevisiae/ultrastructure
6.
J Biol Chem ; 276(18): 15034-41, 2001 May 04.
Article in English | MEDLINE | ID: mdl-11154700

ABSTRACT

We have studied how Pex5p recognizes peroxisomal targeting signal type 1 (PTS1)-containing proteins. A randomly mutagenized pex5 library was screened in a two-hybrid setup for mutations that disrupted the interaction with the PTS1 protein Mdh3p or for suppressor mutations that could restore the interaction with Mdh3p containing a mutation in its PTS1. All mutations localized in the tetratricopeptide repeat (TPR) domain of Pex5p. The Pex5p TPR domain was modeled based on the crystal structure of a related TPR protein. Mapping of the mutations on this structural model revealed that some of the loss-of-interaction mutations consisted of substitutions in alpha-helices of TPRs with bulky amino acids, probably resulting in local misfolding and thereby indirectly preventing binding of PTS1 proteins. The other loss-of-interaction mutations and most suppressor mutations localized in short, exposed, intra-repeat loops of TPR2, TPR3, and TPR6, which are predicted to mediate direct interaction with PTS1 amino acids. Additional site-directed mutants at conserved positions in intra-repeat loops underscored the importance of the loops of TPR2 and TPR3 for PTS1 interaction. Based on the mutational analysis and the structural model, we put forward a model as to how PTS1 proteins are selected by Pex5p.


Subject(s)
Peroxisomes/metabolism , Receptors, Cytoplasmic and Nuclear/metabolism , Amino Acid Sequence , Base Sequence , DNA Primers , Models, Molecular , Molecular Sequence Data , Mutagenesis , Peroxisome-Targeting Signal 1 Receptor , Protein Binding , Protein Conformation , Receptors, Cytoplasmic and Nuclear/chemistry , Receptors, Cytoplasmic and Nuclear/genetics , Sequence Homology, Amino Acid
7.
EMBO J ; 19(23): 6382-91, 2000 Dec 01.
Article in English | MEDLINE | ID: mdl-11101511

ABSTRACT

Src homology 3 (SH3) domains are small non-catalytic protein modules capable of mediating protein-protein interactions by binding to proline-X-X-proline (P-X-X-P) motifs. Here we demonstrate that the SH3 domain of the integral peroxisomal membrane protein Pex13p is able to bind two proteins, one of which, Pex5p, represents a novel non-P-X-X-P ligand. Using alanine scanning, two-hybrid and in vitro interaction analysis, we show that an alpha-helical element in Pex5p is necessary and sufficient for SH3 interaction. Sup pressor analysis using Pex5p mutants located in this alpha-helical element allowed the identification of a unique site of interaction for Pex5p on the Pex13p-SH3 domain that is distinct from the classical P-X-X-P binding pocket. On the basis of a structural model of the Pex13p-SH3 domain we show that this interaction probably takes place between the RT- and distal loops. Thus, the Pex13p-SH3-Pex5p interaction establishes a novel mode of SH3 interaction.


Subject(s)
Membrane Proteins/chemistry , Membrane Proteins/metabolism , Peroxisomes/metabolism , src Homology Domains , Alanine/metabolism , Amino Acid Motifs , Amino Acid Sequence , Binding Sites , Binding, Competitive , Cell Division , Dose-Response Relationship, Drug , Escherichia coli/metabolism , Glutathione Transferase/metabolism , Ligands , Membrane Proteins/genetics , Models, Molecular , Molecular Sequence Data , Mutagenesis, Site-Directed , Peroxisome-Targeting Signal 1 Receptor , Proline/metabolism , Protein Binding , Protein Structure, Secondary , Receptors, Cytoplasmic and Nuclear/chemistry , Receptors, Cytoplasmic and Nuclear/genetics , Receptors, Cytoplasmic and Nuclear/metabolism , Recombinant Fusion Proteins/metabolism , Sequence Homology, Amino Acid , Suppression, Genetic , Two-Hybrid System Techniques
8.
Mol Biol Cell ; 11(11): 3963-76, 2000 Nov.
Article in English | MEDLINE | ID: mdl-11071920

ABSTRACT

A number of peroxisome-associated proteins have been described that are involved in the import of proteins into peroxisomes, among which is the receptor for peroxisomal targeting signal 1 (PTS1) proteins Pex5p, the integral membrane protein Pex13p, which contains an Src homology 3 (SH3) domain, and the peripheral membrane protein Pex14p. In the yeast Saccharomyces cerevisiae, both Pex5p and Pex14p are able to bind Pex13p via its SH3 domain. Pex14p contains the classical SH3 binding motif PXXP, whereas this sequence is absent in Pex5p. Mutation of the conserved tryptophan in the PXXP binding pocket of Pex13-SH3 abolished interaction with Pex14p, but did not affect interaction with Pex5p, suggesting that Pex14p is the classical SH3 domain ligand and that Pex5p binds the SH3 domain in an alternative way. To identify the SH3 binding site in Pex5p, we screened a randomly mutagenized PEX5 library for loss of interaction with Pex13-SH3. Such mutations were all located in a small region in the N-terminal half of Pex5p. One of the altered residues (F208) was part of the sequence W(204)XXQF(208), that is conserved between Pex5 proteins of different species. Site-directed mutagenesis of Trp204 confirmed the essential role of this motif in recognition of the SH3 domain. The Pex5p mutants could only partially restore PTS1-protein import in pex5Delta cells in vivo. In vitro binding studies showed that these Pex5p mutants failed to interact with Pex13-SH3 in the absence of Pex14p, but regained their ability to bind in the presence of Pex14p, suggesting the formation of a heterotrimeric complex consisting of Pex5p, Pex14p, and Pex13-SH3. In vivo, these Pex5p mutants, like wild-type Pex5p, were still found to be associated with peroxisomes. Taken together, this indicates that in the absence of Pex13-SH3 interaction, other protein(s) is able to bind Pex5p at the peroxisome; Pex14p is a likely candidate for this function.


Subject(s)
Membrane Proteins/metabolism , Peroxisomes/metabolism , Receptors, Cytoplasmic and Nuclear/metabolism , Repressor Proteins , Saccharomyces cerevisiae Proteins , Saccharomyces cerevisiae/metabolism , src Homology Domains , Amino Acid Motifs , Amino Acid Sequence , Binding Sites , Carrier Proteins/genetics , Carrier Proteins/metabolism , Membrane Proteins/genetics , Membrane Transport Proteins , Molecular Sequence Data , Mutation , Peroxins , Peroxisome-Targeting Signal 1 Receptor , Protein Transport , Receptors, Cytoplasmic and Nuclear/genetics , Saccharomyces cerevisiae/genetics , Two-Hybrid System Techniques
9.
J Cell Biol ; 150(3): 489-98, 2000 Aug 07.
Article in English | MEDLINE | ID: mdl-10931862

ABSTRACT

The Saccharomyces cerevisiae peroxisomal membrane protein Pex11p has previously been implicated in peroxisome proliferation based on morphological observations of PEX11 mutant cells. Pex11p-deficient cells fail to increase peroxisome number in response to growth on fatty acids and instead accumulate a few giant peroxisomes. We report that mutants deficient in genes required for medium-chain fatty acid (MCFA) beta-oxidation display the same phenotype as Pex11p-deficient cells. Upon closer inspection, we found that Pex11p is required for MCFA beta-oxidation. Disruption of the PEX11 gene results in impaired formation of MCFA-CoA esters as measured in intact cells, whereas their formation is normal in cell lysates. The sole S. cerevisiae MCFA-CoA synthetase (Faa2p) remains properly localized to the inner leaflet of the peroxisomal membrane in PEX11 mutant cells. Therefore, the in vivo latency of MCFA activation observed in Pex11p-deficient cells suggests that Pex11p provides Faa2p with substrate. When PEX11 mutant cells are shifted from glucose to oleate-containing medium, we observed an immediate deficiency in beta-oxidation of MCFAs whereas giant peroxisomes and a failure to increase peroxisome abundance only became apparent much later. Our observations suggest that the MCFA oxidation pathway regulates the level of a signaling molecule that modulates the number of peroxisomal structures in a cell.


Subject(s)
Fatty Acids/metabolism , Membrane Proteins/metabolism , Peroxisomes/physiology , Repressor Proteins , Saccharomyces cerevisiae Proteins , Saccharomyces cerevisiae/metabolism , Acyl-CoA Oxidase , Coenzyme A Ligases/isolation & purification , Coenzyme A Ligases/metabolism , Membrane Proteins/genetics , Mutation , Oleic Acid/metabolism , Oxidation-Reduction , Oxidoreductases/genetics , Peroxins , Peroxisomes/ultrastructure , Saccharomyces cerevisiae/ultrastructure
10.
Biochim Biophys Acta ; 1486(1): 18-27, 2000 Jun 26.
Article in English | MEDLINE | ID: mdl-10856710

ABSTRACT

The peroxisomal membrane forms a permeability barrier for a wide variety of metabolites required for and formed during fatty acid beta-oxidation. To communicate with the cytoplasm and mitochondria, peroxisomes need dedicated proteins to transport such hydrophilic molecules across their membranes. Genetic and biochemical studies in the yeast Saccharomyces cerevisiae have identified enzymes for redox shuttles as well as the first peroxisomal membrane transporter. This peroxisomal ATP-binding cassette transporter (Pat) is highly homologous to the gene mutated in X-linked adrenoleukodystrophy (X-ALD). The yeast Pat is required for import of activated fatty acids into peroxisomes suggesting that this is the primary defect in X-ALD.


Subject(s)
ATP-Binding Cassette Transporters/metabolism , Fatty Acids/metabolism , Intracellular Membranes/metabolism , Peroxisomes/metabolism , ATP Binding Cassette Transporter, Subfamily D, Member 1 , ATP-Binding Cassette Transporters/genetics , Acyl Coenzyme A/metabolism , Biological Transport , Cytoplasm/metabolism , Membrane Proteins/genetics , Oxidation-Reduction , Saccharomyces cerevisiae
13.
EMBO Rep ; 1(1): 40-6, 2000 Jul.
Article in English | MEDLINE | ID: mdl-11256623

ABSTRACT

All eukaryotes so far studied, including animals, plants, yeasts and trypanosomes, have two pathways to target proteins to peroxisomes. These two pathways are specific for the two types of peroxisome targeting signal (PTS) present on peroxisomal matrix proteins. Remarkably, the complete genome sequence of Caenorhabditis elegans lacks the genes encoding proteins specific for the PTS2 targeting pathway. Here we show, by expression of green fluorescent protein (GFP) reporters for both pathways, that the PTS2 pathway is indeed absent in C. elegans. Lack of this pathway in man causes severe disease due to mislocalization of PTS2-containing proteins. This raises the question as to how C. elegans has accommodated the absence of the PTS2 pathway. We found by in silico analysis that C. elegans orthologues of PTS2-containing proteins have acquired a PTS1. We propose that switching of targeting signals has allowed the PTS2 pathway to be lost in the phylogenetic lineage leading to C. elegans.


Subject(s)
Alkyl and Aryl Transferases/metabolism , Caenorhabditis elegans/physiology , Peroxisomes/metabolism , Protein Sorting Signals/genetics , Receptors, Cytoplasmic and Nuclear/metabolism , Alkyl and Aryl Transferases/genetics , Animals , Caenorhabditis elegans/cytology , Caenorhabditis elegans/genetics , Genes, Reporter/genetics , Humans , Microscopy, Fluorescence , Peroxisomal Targeting Signal 2 Receptor , Protein Transport , Recombinant Fusion Proteins/metabolism
14.
Cell Biochem Biophys ; 32 Spring: 1-8, 2000.
Article in English | MEDLINE | ID: mdl-11330035

ABSTRACT

The biogenesis of peroxisomes involves the synthesis of new proteins that after, completion of translation, are targeted to the organelle by virtue of peroxisomal targeting signals (PTS). Two types of PTSs have been well characterized for import of matrix proteins (PTS1 and PTS2). Induction of the genes encoding these matrix proteins takes place in oleate-containing medium and is mediated via an oleate response element (ORE) present in the region preceding these genes. The authors have searched the yeast genome for OREs preceding open reading frames (ORFs), and for ORFs that contain either a PTS1 or PTS2. Of the ORFs containing an ORE, as well as either a PTS1 or a PTS2, many were known to encode bona fide peroxisomal matrix proteins. In addition, candidate genes were identified as encoding putative new peroxisomal proteins. For one case, subcellular location studies validated the in silicio prediction. This gene encodes a new peroxisomal thioesterase.


Subject(s)
Genome, Fungal , Peroxisomes/genetics , Saccharomyces cerevisiae/genetics , Amino Acid Sequence , Molecular Sequence Data , Sequence Alignment , Sequence Analysis, DNA
15.
EMBO J ; 18(21): 5843-52, 1999 Nov 01.
Article in English | MEDLINE | ID: mdl-10545096

ABSTRACT

In Saccharomyces cerevisiae, beta-oxidation of fatty acids is confined to peroxisomes. The acetyl-CoA produced has to be transported from the peroxisomes via the cytoplasm to the mitochondrial matrix in order to be degraded to CO(2) and H(2)O. Two pathways for the transport of acetyl-CoA to the mitochondria have been proposed. The first involves peroxisomal conversion of acetyl-CoA into glyoxylate cycle intermediates followed by transport of these intermediates to the mitochondria. The second pathway involves peroxisomal conversion of acetyl-CoA into acetylcarnitine, which is subsequently transported to the mitochondria. Using a selective screen, we have isolated several mutants that are specifically affected in the second pathway, the carnitine-dependent acetyl-CoA transport from the peroxisomes to the mitochondria, and assigned these CDAT mutants to three different complementation groups. The corresponding genes were identified using functional complementation of the mutants with a genomic DNA library. In addition to the previously reported carnitine acetyl-CoA transferase (CAT2), we identified the genes for the yeast orthologue of the human mitochondrial carnitine acylcarnitine translocase (YOR100C or CAC) and for a transport protein (AGP2) required for carnitine transport across the plasma membrane.


Subject(s)
Acetyl Coenzyme A/metabolism , Amino Acid Transport Systems , Carnitine/metabolism , Carrier Proteins/genetics , Cell Membrane/chemistry , Membrane Proteins/genetics , Mitochondria/metabolism , Peroxisomes/metabolism , Saccharomyces cerevisiae Proteins , Saccharomyces cerevisiae/genetics , Symporters , Biological Transport/genetics , Carnitine Acyltransferases/metabolism , Carnitine O-Acetyltransferase/genetics , Carnitine O-Acetyltransferase/metabolism , Carrier Proteins/metabolism , Cloning, Molecular , Fungal Proteins , Genes, Fungal , Membrane Proteins/chemistry , Microscopy, Immunoelectron , Mutation , Oleic Acid/metabolism , Saccharomyces cerevisiae/metabolism
16.
Trends Cell Biol ; 9(11): 447-53, 1999 Nov.
Article in English | MEDLINE | ID: mdl-10511709

ABSTRACT

Peroxisomes compartmentalize part of the anabolic and catabolic pathways and reactions of the cell. Dysfunction of a single peroxisomal enzyme or loss of the whole peroxisomal compartment causes sporadic, but serious, human diseases. Genetic studies in various yeasts have identified PEX genes, which are required for the maintenance of complete peroxisomes. Mutations in PEX genes have proved to be the molecular cause of several human diseases, particularly those involving loss of organelles. Peroxisomes have several properties that distinguish them from other organelles, including the import of folded proteins from the cytosol by an unknown mechanism. By discussing recent highlights from the field of peroxisome research, we aim to share with the general readership our excitement as well as the many mysteries still surrounding peroxisome function and maintenance.


Subject(s)
Peroxisomes/metabolism , Animals , Biological Transport , Humans , Peroxisomes/genetics , Protein Folding , Proteins/metabolism , Rats
17.
Biochim Biophys Acta ; 1451(1): 17-34, 1999 Aug 12.
Article in English | MEDLINE | ID: mdl-10446385

ABSTRACT

Peroxisomes are organelles that confine an important set of enzymes within their single membrane boundaries. In man, a wide variety of genetic disorders is caused by loss of peroxisome function. In the most severe cases, the clinical phenotype indicates that abnormalities begin to appear during embryological development. In less severe cases, the quality of life of adults is affected. Research on yeast model systems has contributed to a better understanding of peroxisome formation and maintenance. This framework of knowledge has made it possible to understand the molecular basis of most of the peroxisome biogenesis disorders. Interestingly, most peroxisome biogenesis disorders are caused by a failure to target peroxisomal proteins to the organellar matrix or membrane, which classifies them as protein targeting diseases. Here we review recent fundamental research on peroxisomal protein targeting and discuss a few burning questions in the field concerning the origin of peroxisomes.


Subject(s)
Intracellular Membranes/chemistry , Microbodies/chemistry , Proteins/chemistry , Membrane Proteins/chemistry , Peroxisomal Targeting Signal 2 Receptor , Peroxisome-Targeting Signal 1 Receptor , Protein Folding , Receptors, Cytoplasmic and Nuclear/chemistry , Signal Transduction
18.
FEBS Lett ; 453(1-2): 210-4, 1999 Jun 18.
Article in English | MEDLINE | ID: mdl-10403405

ABSTRACT

Conditions that stress the endoplasmic reticulum (ER) in Saccharomyces cerevisiae can elicit a combination of an unfolded protein response (UPR) and an inositol response (IR). This results in increased synthesis of ER protein-folding factors and of enzymes participating in phospholipid biosynthesis. It was suggested that in cells grown on glucose or galactose medium, the UPR and the IR are linked and controlled by the ER stress sensor Ire1p. However, our studies suggest that during growth on oleate the IR is controlled both by an Ire1p-dependent pathway and by an Ire1p-independent pathway.


Subject(s)
Endoplasmic Reticulum/physiology , Fungal Proteins/metabolism , Membrane Glycoproteins/metabolism , Microbodies/physiology , Protein Folding , Protein Serine-Threonine Kinases , Saccharomyces cerevisiae Proteins , Saccharomyces cerevisiae/physiology , Adaptation, Biological , Endoplasmic Reticulum/ultrastructure , Galactose/metabolism , Gene Deletion , Membrane Proteins/biosynthesis , Myo-Inositol-1-Phosphate Synthase/biosynthesis , Oleic Acid/metabolism , Phosphoproteins/biosynthesis
20.
Mol Biol Cell ; 10(6): 1859-72, 1999 Jun.
Article in English | MEDLINE | ID: mdl-10359602

ABSTRACT

We describe a genome-wide characterization of mRNA transcript levels in yeast grown on the fatty acid oleate, determined using Serial Analysis of Gene Expression (SAGE). Comparison of this SAGE library with that reported for glucose grown cells revealed the dramatic adaptive response of yeast to a change in carbon source. A major fraction (>20%) of the 15,000 mRNA molecules in a yeast cell comprised differentially expressed transcripts, which were derived from only 2% of the total number of approximately 6300 yeast genes. Most of the mRNAs that were differentially expressed code for enzymes or for other proteins participating in metabolism (e.g., metabolite transporters). In oleate-grown cells, this was exemplified by the huge increase of mRNAs encoding the peroxisomal beta-oxidation enzymes required for degradation of fatty acids. The data provide evidence for the existence of redox shuttles across organellar membranes that involve peroxisomal, cytoplasmic, and mitochondrial enzymes. We also analyzed the mRNA profile of a mutant strain with deletions of the PIP2 and OAF1 genes, encoding transcription factors required for induction of genes encoding peroxisomal proteins. Induction of genes under the immediate control of these factors was abolished; other genes were up-regulated, indicating an adaptive response to the changed metabolism imposed by the genetic impairment. We describe a statistical method for analysis of data obtained by SAGE.


Subject(s)
Carbon/metabolism , Gene Expression Regulation, Fungal , Saccharomyces cerevisiae Proteins , Saccharomyces cerevisiae/genetics , Saccharomyces cerevisiae/metabolism , Cytosol/metabolism , Fungal Proteins/genetics , Fungal Proteins/metabolism , Gene Library , Genetic Techniques , Glucose/metabolism , Heat-Shock Proteins/genetics , Heat-Shock Proteins/metabolism , Microbodies/genetics , Microbodies/metabolism , Mitochondria/genetics , Mitochondria/metabolism , Models, Statistical , Mutation , Oleic Acid/metabolism , Transcription Factors/genetics , Transcription Factors/metabolism , Transcription, Genetic
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