Identification of a Peroxisomal ATP Carrier Required for Medium-Chain Fatty Acid β-Oxidation and Normal Peroxisome Proliferation in Saccharomyces cerevisiae

ABSTRACT We have characterized the role of YPR128cp, the orthologue of human PMP34, in fatty acid metabolism and peroxisomal proliferation inSaccharomyces cerevisiae. YPR128cp belongs to the mitochondrial carrier family (MCF) of solute transporters and is localized in the peroxisomal membrane. Disruption of theYPR128c 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 β-oxidation activity was markedly reduced in intact ypr128cΔ 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 β-oxidation is impaired in ypr128cΔ cells. Since MCFA β-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 ypr128cΔ 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.

[1]  H. Tabak,et al.  Pex11p Plays a Primary Role in Medium-Chain Fatty Acid Oxidation, a Process That Affects Peroxisome Number and Size in Saccharomyces cerevisiae , 2000, The Journal of cell biology.

[2]  D. Valle,et al.  Peroxisome biogenesis disorders: genetics and cell biology. , 2000, Trends in genetics : TIG.

[3]  A. Yamashita,et al.  Peroxisomal Membrane Protein Pmp47 Is Essential in the Metabolism of Middle-chain Fatty Acid in Yeast Peroxisomes and Is Associated with Peroxisome Proliferation* , 2000, The Journal of Biological Chemistry.

[4]  F. Kamp,et al.  How are free fatty acids transported in membranes? Is it by proteins or by free diffusion through the lipids? , 1999, Diabetes.

[5]  H. Tabak,et al.  Peroxisomes: simple in function but complex in maintenance. , 1999, Trends in cell biology.

[6]  H. Tabak,et al.  Molecular characterization of carnitine‐dependent transport of acetyl‐CoA from peroxisomes to mitochondria in Saccharomyces cerevisiae and identification of a plasma membrane carnitine transporter, Agp2p , 1999, The EMBO journal.

[7]  H. Kennedy,et al.  Glucose Generates Sub-plasma Membrane ATP Microdomains in Single Islet β-Cells , 1999, The Journal of Biological Chemistry.

[8]  H. Kennedy,et al.  Glucose generates sub-plasma membrane ATP microdomains in single islet beta-cells. Potential role for strategically located mitochondria. , 1999, The Journal of biological chemistry.

[9]  P. Hieter,et al.  Detecting patterns of protein distribution and gene expression in silico. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[10]  C. Brees,et al.  Identification and characterization of human PMP34, a protein closely related to the peroxisomal integral membrane protein PMP47 of Candida boidinii. , 1998, European journal of biochemistry.

[11]  Igor V. Karpichev,et al.  Global Regulatory Functions of Oaf1p and Pip2p (Oaf2p), Transcription Factors That Regulate Genes Encoding Peroxisomal Proteins in Saccharomyces cerevisiae , 1998, Molecular and Cellular Biology.

[12]  V. Iacobazzi,et al.  Bacterial overexpression, purification, and reconstitution of the carnitine/acylcarnitine carrier from rat liver mitochondria. , 1998, Biochemical and biophysical research communications.

[13]  M H Saier,et al.  Unified inventory of established and putative transporters encoded within the complete genome of Saccharomyces cerevisiae , 1998, FEBS letters.

[14]  H. Tabak,et al.  Peroxisomal β‐oxidation of polyunsaturated fatty acids in Saccharomyces cerevisiae: isocitrate dehydrogenase provides NADPH for reduction of double bonds at even positions , 1998, The EMBO journal.

[15]  S. Subramani.  Components involved in peroxisome import, biogenesis, proliferation, turnover, and movement. , 1998, Physiological reviews.

[16]  H. Tabak,et al.  Transport of activated fatty acids by the peroxisomal ATP-binding-cassette transporter Pxa2 in a semi-intact yeast cell system. , 1997, European journal of biochemistry.

[17]  J. Lamotte‐Brasseur,et al.  Phylogenetic Classification of the Mitochondrial Carrier Family of Saccharomyces cerevisiae , 1997, Yeast.

[18]  D. Kakhniashvili,et al.  Identification of a Novel Gene Encoding the Yeast Mitochondrial Dicarboxylate Transport Protein via Overexpression, Purification, and Characterization of Its Protein Product* , 1997, The Journal of Biological Chemistry.

[19]  H. Tabak,et al.  Analysis of the Carboxyl-terminal Peroxisomal Targeting Signal 1 in a Homologous Context in Saccharomyces cerevisiae* , 1996, The Journal of Biological Chemistry.

[20]  D. Valle,et al.  A Saccharomyces cerevisiae homolog of the human adrenoleukodystrophy transporter is a heterodimer of two half ATP-binding cassette transporters. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[21]  M. De Rijcke,et al.  The ARG11 Gene of Saccharomyces cerevisiae Encodes a Mitochondrial Integral Membrane Protein Required for Arginine Biosynthesis* , 1996, The Journal of Biological Chemistry.

[22]  J. Dyer,et al.  Redox-sensitive homodimerization of Pex11p: a proposed mechanism to regulate peroxisomal division , 1996, The Journal of cell biology.

[23]  C. Rodrigues-Pousada,et al.  The ABC transporter proteins Pat1 and Pat2 are required for import of long‐chain fatty acids into peroxisomes of Saccharomyces cerevisiae. , 1996, The EMBO journal.

[24]  H. Saiki,et al.  The absence of Pmp47, a putative yeast peroxisomal transporter, causes a defect in transport and folding of a specific matrix enzyme , 1996, The Journal of cell biology.

[25]  H. Tabak,et al.  Pip2p: a transcriptional regulator of peroxisome proliferation in the yeast Saccharomyces cerevisiae. , 1996, The EMBO journal.

[26]  A. Wach.  PCR‐synthesis of marker cassettes with long flanking homology regions for gene disruptions in S. cerevisiae , 1996, Yeast.

[27]  J. Thorner,et al.  The PAL1 gene product is a peroxisomal ATP-binding cassette transporter in the yeast Saccharomyces cerevisiae , 1996, The Journal of cell biology.

[28]  H. Schulz,et al.  beta-oxidation of fatty acids in mitochondria, peroxisomes, and bacteria: a century of continued progress. , 1995, Progress in lipid research.

[29]  S. R. Ford,et al.  Does the sulfhydryl or the adenine moiety of CoA enhance firefly luciferase activity? , 1995, Biochimica et biophysica acta.

[30]  H. Tabak,et al.  The membrane of peroxisomes in Saccharomyces cerevisiae is impermeable to NAD(H) and acetyl‐CoA under in vivo conditions. , 1995, The EMBO journal.

[31]  D. Valle,et al.  PXA1, a possible Saccharomyces cerevisiae ortholog of the human adrenoleukodystrophy gene. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[32]  J. Dyer,et al.  Pmp27 promotes peroxisomal proliferation , 1995, The Journal of cell biology.

[33]  D. Wood,et al.  High Level Expression and Characterization of the Mitochondrial Citrate Transport Protein from the Yeast Saccharomyces cerevisiae(*) , 1995, The Journal of Biological Chemistry.

[34]  P. M. Sokolove,et al.  The reconstituted mitochondrial adenine nucleotide translocator: effects of lipid polymorphism. , 1994, Archives of biochemistry and biophysics.

[35]  H. Feldmann,et al.  II. Yeast sequencing reports. Analysis of a 70kb region on the right arm of yeast chromosome II , 1994 .

[36]  C. Nombela,et al.  Expression and in vivo determination of firefly luciferase as gene reporter in Saccharomyces cerevisiae , 1994, Yeast.

[37]  H. Feldmann,et al.  Analysis of a 70 kb region on the right arm of yeast chromosome II. , 1994, Yeast.

[38]  J. Reddy,et al.  Peroxisomal lipid metabolism. , 1994, Annual review of nutrition.

[39]  G. Braus,et al.  YMC1, a yeast gene encoding a new putative mitochondrial carrier protein , 1993, Yeast.

[40]  A. Munnich,et al.  Human trifunctional protein deficiency: a new disorder of mitochondrial fatty acid beta-oxidation. , 1992, Biochemical and biophysical research communications.

[41]  H. Tabak,et al.  Isolation of peroxisome assembly mutants from Saccharomyces cerevisiae with different morphologies using a novel positive selection procedure , 1992, The Journal of cell biology.

[42]  T. Baldwin,et al.  Use of bacterial and firefly luciferases as reporter genes in DEAE-dextran-mediated transfection of mammalian cells. , 1992, Analytical biochemistry.

[43]  B. Dujon,et al.  Sequence of a segment of yeast chromosome XI identifies a new mitochondrial carrier,a new member of the G protein family, and a protein with the PAAKK motif of the H1 histones , 1992, Yeast.

[44]  R. Wanders,et al.  Biochemistry of peroxisomes. , 1992, Annual review of biochemistry.

[45]  C. Schobert,et al.  Cloning and characterization of the mitochondrial phosphate transport protein gene from the yeast Saccharomyces cerevisiae. , 1991, Biochemistry.

[46]  R. Schweyen,et al.  MRS3 and MRS4, two suppressors of mtRNA splicing defects in yeast, are new members of the mitochondrial carrier family. , 1991, Journal of molecular biology.

[47]  C. Slaughter,et al.  Sorting of peroxisomal membrane protein PMP47 from Candida boidinii into peroxisomal membranes of Saccharomyces cerevisiae. , 1990, The Journal of biological chemistry.

[48]  N. Nelson,et al.  A third ADP/ATP translocator gene in yeast. , 1990, The Journal of biological chemistry.

[49]  H. Tabak,et al.  Peroxisomal protein import is conserved between yeast, plants, insects and mammals. , 1990, The EMBO journal.

[50]  J. E. Lawson,et al.  Separate genes encode functionally equivalent ADP/ATP carrier proteins in Saccharomyces cerevisiae. Isolation and analysis of AAC2. , 1988, The Journal of biological chemistry.

[51]  N. Gordon.  Peroxisomal disorders , 1987, Brain and Development.

[52]  D. Montgomery,et al.  Sequences required for delivery and localization of the ADP/ATP translocator to the mitochondrial inner membrane , 1986, Molecular and cellular biology.

[53]  P. K. Smith,et al.  Measurement of protein using bicinchoninic acid. , 1985, Analytical biochemistry.

[54]  S. Akira,et al.  Immunoglobulin gene expression and DNA methylation in murine pre‐B cell lines. , 1984, The EMBO journal.