Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 2002 Jul 1;365(Pt 1):109–117. doi: 10.1042/BJ20011495

The peroxisomal transporter gene ANT1 is regulated by a deviant oleate response element (ORE): characterization of the signal for fatty acid induction.

Hanspeter Rottensteiner 1, Luigi Palmieri 1, Andreas Hartig 1, Barbara Hamilton 1, Helmut Ruis 1, Ralf Erdmann 1, Aner Gurvitz 1
PMCID: PMC1222661  PMID: 12071844

Abstract

Saccharomyces cerevisiae ANT1/YPR128c encodes the peroxisomal adenine nucleotide transporter that provides ATP for intra-peroxisomal activation of medium-chain fatty acids. A lacZ reporter construct comprising the ANT1 promoter was shown to be comparatively more highly expressed in a wild-type strain grown on oleic acid, a long-chain fatty acid, than in pip2Delta(oaf1)Delta mutant cells that are defective in fatty acid induction. The ANT1 promoter was demonstrated to contain a deviant oleate response element (ORE) that could bind the Pip2p-Oaf1p transcription factor and confer activation on a basal CYC1-lacZ reporter gene. Expression of Ant1p as well as other enzymes whose genes are known to be regulated by a canonical ORE was found to be increased in cells grown on lauric acid, a medium-chain fatty acid. We concluded that the signal for induction does not differentiate between long- and medium-chain fatty acids. This signal was independent of beta-oxidation or the biogenesis of the peroxisomal compartment where this process occurs, since a pox1Delta strain blocked in the first and rate-limiting step of beta-oxidation as well as various pex mutant cells devoid of intact peroxisomes produced sufficient amounts of Pip2p-Oaf1p for binding OREs in vitro and for expressing an ORE-driven reporter gene. The signal's durability was shown to be related to the concentration of fatty acids in the medium, since a pex6Delta strain expressed an ORE-driven reporter gene at high levels for a longer period than did isogenic wild-type cells. Generation of the signal was also independent of protein synthesis, as demonstrated by cycloheximide treatment.

Full Text

The Full Text of this article is available as a PDF (238.1 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. BEERS R. F., Jr, SIZER I. W. A spectrophotometric method for measuring the breakdown of hydrogen peroxide by catalase. J Biol Chem. 1952 Mar;195(1):133–140. [PubMed] [Google Scholar]
  2. Baumgartner U., Hamilton B., Piskacek M., Ruis H., Rottensteiner H. Functional analysis of the Zn(2)Cys(6) transcription factors Oaf1p and Pip2p. Different roles in fatty acid induction of beta-oxidation in Saccharomyces cerevisiae. J Biol Chem. 1999 Aug 6;274(32):22208–22216. doi: 10.1074/jbc.274.32.22208. [DOI] [PubMed] [Google Scholar]
  3. Brachmann C. B., Davies A., Cost G. J., Caputo E., Li J., Hieter P., Boeke J. D. Designer deletion strains derived from Saccharomyces cerevisiae S288C: a useful set of strains and plasmids for PCR-mediated gene disruption and other applications. Yeast. 1998 Jan 30;14(2):115–132. doi: 10.1002/(SICI)1097-0061(19980130)14:2<115::AID-YEA204>3.0.CO;2-2. [DOI] [PubMed] [Google Scholar]
  4. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  5. Chen D. C., Yang B. C., Kuo T. T. One-step transformation of yeast in stationary phase. Curr Genet. 1992 Jan;21(1):83–84. doi: 10.1007/BF00318659. [DOI] [PubMed] [Google Scholar]
  6. Dmochowska A., Dignard D., Maleszka R., Thomas D. Y. Structure and transcriptional control of the Saccharomyces cerevisiae POX1 gene encoding acyl-coenzyme A oxidase. Gene. 1990 Apr 16;88(2):247–252. doi: 10.1016/0378-1119(90)90038-s. [DOI] [PubMed] [Google Scholar]
  7. Einerhand A. W., Kos W. T., Distel B., Tabak H. F. Characterization of a transcriptional control element involved in proliferation of peroxisomes in yeast in response to oleate. Eur J Biochem. 1993 May 15;214(1):323–331. doi: 10.1111/j.1432-1033.1993.tb17927.x. [DOI] [PubMed] [Google Scholar]
  8. Erdmann R., Veenhuis M., Mertens D., Kunau W. H. Isolation of peroxisome-deficient mutants of Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1989 Jul;86(14):5419–5423. doi: 10.1073/pnas.86.14.5419. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Filipits M., Simon M. M., Rapatz W., Hamilton B., Ruis H. A Saccharomyces cerevisiae upstream activating sequence mediates induction of peroxisome proliferation by fatty acids. Gene. 1993 Sep 30;132(1):49–55. doi: 10.1016/0378-1119(93)90513-3. [DOI] [PubMed] [Google Scholar]
  10. Forman B. M., Chen J., Evans R. M. Hypolipidemic drugs, polyunsaturated fatty acids, and eicosanoids are ligands for peroxisome proliferator-activated receptors alpha and delta. Proc Natl Acad Sci U S A. 1997 Apr 29;94(9):4312–4317. doi: 10.1073/pnas.94.9.4312. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Geisbrecht B. V., Schulz K., Nau K., Geraghty M. T., Schulz H., Erdmann R., Gould S. J. Preliminary characterization of Yor180Cp: identification of a novel peroxisomal protein of saccharomyces cerevisiae involved in fatty acid metabolism. Biochem Biophys Res Commun. 1999 Jun 24;260(1):28–34. doi: 10.1006/bbrc.1999.0860. [DOI] [PubMed] [Google Scholar]
  12. Geisbrecht B. V., Zhu D., Schulz K., Nau K., Morrell J. C., Geraghty M., Schulz H., Erdmann R., Gould S. J. Molecular characterization of Saccharomyces cerevisiae Delta3, Delta2-enoyl-CoA isomerase. J Biol Chem. 1998 Dec 11;273(50):33184–33191. doi: 10.1074/jbc.273.50.33184. [DOI] [PubMed] [Google Scholar]
  13. Geraghty M. T., Bassett D., Morrell J. C., Gatto G. J., Jr, Bai J., Geisbrecht B. V., Hieter P., Gould S. J. Detecting patterns of protein distribution and gene expression in silico. Proc Natl Acad Sci U S A. 1999 Mar 16;96(6):2937–2942. doi: 10.1073/pnas.96.6.2937. [DOI] [PMC free article] [PubMed] [Google Scholar] [Retracted]
  14. Gurvitz A., Hamilton B., Hartig A., Ruis H., Dawes I. W., Rottensteiner H. A novel element in the promoter of the Saccharomyces cerevisiae gene SPS19 enhances ORE-dependent up-regulation in oleic acid and is essential for de-repression. Mol Gen Genet. 1999 Oct;262(3):481–492. doi: 10.1007/s004380051109. [DOI] [PubMed] [Google Scholar]
  15. Gurvitz A., Hamilton B., Ruis H., Hartig A. Peroxisomal degradation of trans-unsaturated fatty acids in the yeast Saccharomyces cerevisiae. J Biol Chem. 2001 Jan 12;276(2):895–903. doi: 10.1074/jbc.M003305200. [DOI] [PubMed] [Google Scholar]
  16. Gurvitz A., Hiltunen J. K., Erdmann R., Hamilton B., Hartig A., Ruis H., Rottensteiner H. Saccharomyces cerevisiae Adr1p governs fatty acid beta-oxidation and peroxisome proliferation by regulating POX1 and PEX11. J Biol Chem. 2001 Jun 28;276(34):31825–31830. doi: 10.1074/jbc.M105989200. [DOI] [PubMed] [Google Scholar]
  17. Gurvitz A., Mursula A. M., Firzinger A., Hamilton B., Kilpeläinen S. H., Hartig A., Ruis H., Hiltunen J. K., Rottensteiner H. Peroxisomal Delta3-cis-Delta2-trans-enoyl-CoA isomerase encoded by ECI1 is required for growth of the yeast Saccharomyces cerevisiae on unsaturated fatty acids. J Biol Chem. 1998 Nov 20;273(47):31366–31374. doi: 10.1074/jbc.273.47.31366. [DOI] [PubMed] [Google Scholar]
  18. Gurvitz A., Mursula A. M., Yagi A. I., Hartig A., Ruis H., Rottensteiner H., Hiltunen J. K. Alternatives to the isomerase-dependent pathway for the beta-oxidation of oleic acid are dispensable in Saccharomyces cerevisiae. Identification of YOR180c/DCI1 encoding peroxisomal delta(3,5)-delta(2,4)-dienoyl-CoA isomerase. J Biol Chem. 1999 Aug 27;274(35):24514–24521. doi: 10.1074/jbc.274.35.24514. [DOI] [PubMed] [Google Scholar]
  19. Gurvitz A., Rottensteiner H., Hiltunen J. K., Binder M., Dawes I. W., Ruis H., Hamilton B. Regulation of the yeast SPS19 gene encoding peroxisomal 2,4-dienoyl-CoA reductase by the transcription factors Pip2p and Oaf1p: beta-oxidation is dispensable for Saccharomyces cerevisiae sporulation in acetate medium. Mol Microbiol. 1997 Nov;26(4):675–685. doi: 10.1046/j.1365-2958.1997.5931969.x. [DOI] [PubMed] [Google Scholar]
  20. Gurvitz A., Rottensteiner H., Kilpeläinen S. H., Hartig A., Hiltunen J. K., Binder M., Dawes I. W., Hamilton B. The Saccharomyces cerevisiae peroxisomal 2,4-dienoyl-CoA reductase is encoded by the oleate-inducible gene SPS19. J Biol Chem. 1997 Aug 29;272(35):22140–22147. doi: 10.1074/jbc.272.35.22140. [DOI] [PubMed] [Google Scholar]
  21. Gurvitz A., Wabnegger L., Rottensteiner H., Dawes I. W., Hartig A., Ruis H., Hamilton B. Adr1p-dependent regulation of the oleic acid-inducible yeast gene SPS19 encoding the peroxisomal beta-oxidation auxiliary enzyme 2,4-dienoyl-CoA reductase. Mol Cell Biol Res Commun. 2000 Aug;4(2):81–89. doi: 10.1006/mcbr.2000.0261. [DOI] [PubMed] [Google Scholar]
  22. Hettema E. H., van Roermund C. W., Distel B., van den Berg M., Vilela C., Rodrigues-Pousada C., Wanders R. J., Tabak H. F. The ABC transporter proteins Pat1 and Pat2 are required for import of long-chain fatty acids into peroxisomes of Saccharomyces cerevisiae. EMBO J. 1996 Aug 1;15(15):3813–3822. [PMC free article] [PubMed] [Google Scholar]
  23. Igual J. C., Navarro B. Respiration and low cAMP-dependent protein kinase activity are required for high-level expression of the peroxisomal thiolase gene in Saccharomyces cerevisiae. Mol Gen Genet. 1996 Sep 25;252(4):446–455. doi: 10.1007/BF02173010. [DOI] [PubMed] [Google Scholar]
  24. Jones E. W. Proteinase mutants of Saccharomyces cerevisiae. Genetics. 1977 Jan;85(1):23–33. doi: 10.1093/genetics/85.1.23. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Kal A. J., van Zonneveld A. J., Benes V., van den Berg M., Koerkamp M. G., Albermann K., Strack N., Ruijter J. M., Richter A., Dujon B. Dynamics of gene expression revealed by comparison of serial analysis of gene expression transcript profiles from yeast grown on two different carbon sources. Mol Biol Cell. 1999 Jun;10(6):1859–1872. doi: 10.1091/mbc.10.6.1859. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Karpichev I. V., Luo Y., Marians R. C., Small G. M. A complex containing two transcription factors regulates peroxisome proliferation and the coordinate induction of beta-oxidation enzymes in Saccharomyces cerevisiae. Mol Cell Biol. 1997 Jan;17(1):69–80. doi: 10.1128/mcb.17.1.69. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Karpichev I. V., Small G. M. Global regulatory functions of Oaf1p and Pip2p (Oaf2p), transcription factors that regulate genes encoding peroxisomal proteins in Saccharomyces cerevisiae. Mol Cell Biol. 1998 Nov;18(11):6560–6570. doi: 10.1128/mcb.18.11.6560. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Kliewer S. A., Sundseth S. S., Jones S. A., Brown P. J., Wisely G. B., Koble C. S., Devchand P., Wahli W., Willson T. M., Lenhard J. M. Fatty acids and eicosanoids regulate gene expression through direct interactions with peroxisome proliferator-activated receptors alpha and gamma. Proc Natl Acad Sci U S A. 1997 Apr 29;94(9):4318–4323. doi: 10.1073/pnas.94.9.4318. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Kunau W. H., Dommes V., Schulz H. beta-oxidation of fatty acids in mitochondria, peroxisomes, and bacteria: a century of continued progress. Prog Lipid Res. 1995;34(4):267–342. doi: 10.1016/0163-7827(95)00011-9. [DOI] [PubMed] [Google Scholar]
  30. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  31. Luo Y., Karpichev I. V., Kohanski R. A., Small G. M. Purification, identification, and properties of a Saccharomyces cerevisiae oleate-activated upstream activating sequence-binding protein that is involved in the activation of POX1. J Biol Chem. 1996 May 17;271(20):12068–12075. doi: 10.1074/jbc.271.20.12068. [DOI] [PubMed] [Google Scholar]
  32. Marzioch M., Erdmann R., Veenhuis M., Kunau W. H. PAS7 encodes a novel yeast member of the WD-40 protein family essential for import of 3-oxoacyl-CoA thiolase, a PTS2-containing protein, into peroxisomes. EMBO J. 1994 Oct 17;13(20):4908–4918. doi: 10.1002/j.1460-2075.1994.tb06818.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Myers A. M., Tzagoloff A., Kinney D. M., Lusty C. J. Yeast shuttle and integrative vectors with multiple cloning sites suitable for construction of lacZ fusions. Gene. 1986;45(3):299–310. doi: 10.1016/0378-1119(86)90028-4. [DOI] [PubMed] [Google Scholar]
  34. Palmieri L., Rottensteiner H., Girzalsky W., Scarcia P., Palmieri F., Erdmann R. Identification and functional reconstitution of the yeast peroxisomal adenine nucleotide transporter. EMBO J. 2001 Sep 17;20(18):5049–5059. doi: 10.1093/emboj/20.18.5049. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Richter K., Ammerer G., Hartter E., Ruis H. The effect of delta-aminolevulinate on catalase T-messenger RNA levels in delta-aminolevulinate synthase-defective mutants of Saccharomyces cerevisiae. J Biol Chem. 1980 Sep 10;255(17):8019–8022. [PubMed] [Google Scholar]
  36. Rottensteiner H., Kal A. J., Filipits M., Binder M., Hamilton B., Tabak H. F., Ruis H. Pip2p: a transcriptional regulator of peroxisome proliferation in the yeast Saccharomyces cerevisiae. EMBO J. 1996 Jun 17;15(12):2924–2934. [PMC free article] [PubMed] [Google Scholar]
  37. Rottensteiner H., Kal A. J., Hamilton B., Ruis H., Tabak H. F. A heterodimer of the Zn2Cys6 transcription factors Pip2p and Oaf1p controls induction of genes encoding peroxisomal proteins in Saccharomyces cerevisiae. Eur J Biochem. 1997 Aug 1;247(3):776–783. doi: 10.1111/j.1432-1033.1997.00776.x. [DOI] [PubMed] [Google Scholar]
  38. Shani N., Valle D. A Saccharomyces cerevisiae homolog of the human adrenoleukodystrophy transporter is a heterodimer of two half ATP-binding cassette transporters. Proc Natl Acad Sci U S A. 1996 Oct 15;93(21):11901–11906. doi: 10.1073/pnas.93.21.11901. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Simon M., Adam G., Rapatz W., Spevak W., Ruis H. The Saccharomyces cerevisiae ADR1 gene is a positive regulator of transcription of genes encoding peroxisomal proteins. Mol Cell Biol. 1991 Feb;11(2):699–704. doi: 10.1128/mcb.11.2.699. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Skoneczny M., Chełstowska A., Rytka J. Study of the coinduction by fatty acids of catalase A and acyl-CoA oxidase in standard and mutant Saccharomyces cerevisiae strains. Eur J Biochem. 1988 Jun 1;174(2):297–302. doi: 10.1111/j.1432-1033.1988.tb14097.x. [DOI] [PubMed] [Google Scholar]
  41. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  42. Van der Leij I., Franse M. M., Elgersma Y., Distel B., Tabak H. F. PAS10 is a tetratricopeptide-repeat protein that is essential for the import of most matrix proteins into peroxisomes of Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1993 Dec 15;90(24):11782–11786. doi: 10.1073/pnas.90.24.11782. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Veenhuis M., Mateblowski M., Kunau W. H., Harder W. Proliferation of microbodies in Saccharomyces cerevisiae. Yeast. 1987 Jun;3(2):77–84. doi: 10.1002/yea.320030204. [DOI] [PubMed] [Google Scholar]
  44. Voorn-Brouwer T., van der Leij I., Hemrika W., Distel B., Tabak H. F. Sequence of the PAS8 gene, the product of which is essential for biogenesis of peroxisomes in Saccharomyces cerevisiae. Biochim Biophys Acta. 1993 Nov 16;1216(2):325–328. doi: 10.1016/0167-4781(93)90166-b. [DOI] [PubMed] [Google Scholar]
  45. Yaffe M. P., Schatz G. Two nuclear mutations that block mitochondrial protein import in yeast. Proc Natl Acad Sci U S A. 1984 Aug;81(15):4819–4823. doi: 10.1073/pnas.81.15.4819. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. van Roermund C. W., Drissen R., van Den Berg M., Ijlst L., Hettema E. H., Tabak H. F., Waterham H. R., Wanders R. J. Identification of a peroxisomal ATP carrier required for medium-chain fatty acid beta-oxidation and normal peroxisome proliferation in Saccharomyces cerevisiae. Mol Cell Biol. 2001 Jul;21(13):4321–4329. doi: 10.1128/MCB.21.13.4321-4329.2001. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES