Advertisement

Archives of Microbiology

, Volume 201, Issue 1, pp 123–134 | Cite as

Regulation of the inositol transporter Itr1p by hydrogen peroxide in Saccharomyces cerevisiae

  • Tomás Santos
  • H. Susana Marinho
  • Luisa Cyrne
Original Paper
  • 81 Downloads

Abstract

Myo-inositol is a precursor of several membrane phospholipids and sphingolipids and plays a key role in gene regulation in Saccharomyces cerevisiae (S. cerevisiae). Here, we tested whether H2O2 was affecting the levels of the inositol transporters and thus inositol uptake. In S. cerevisiae cells adapted to H2O2 Itr1–GFPp accumulated in the plasma membrane until 20 min, concomitantly with an inhibition of its internalization. Exposure to H2O2 did not alter Itr2–GFPp cellular levels and induced only an 8% decrease at 10 min in the plasma membrane. Therefore, decreased inositol intracellular levels are not caused by decreased levels of inositol transporters in the plasma membrane. However, results show that H2O2 adaptation affects Itr1p turnover and, consequently, H2O2-adapted yeast cells display an inositol transporter phenotype comparable to cells grown in the absence of inositol in growth medium, i.e. accumulation in the plasma membrane and decreased degradation.

Keywords

Redox signaling Hydrogen peroxide Saccharomyces cerevisiae Myo-inositol transporters Inositol Adaptation 

Abbreviations

ANOVA

One-way analysis of variance

Art5p

Arrestin-related trafficking adaptors

GFP–Opi1p

Fusion between green fluorescence protein and Opi1 protein

H2O2

Hydrogen peroxide

INO1

Inositol-1-phosphate synthase gene

INO2

Ino2p gene

ITR1

Myo-inositol transporter 1 gene

Itr1–GFPp

Fusion between Itr1 protein and green fluorescence protein

ITR2

Myo-inositol transporter 2 gene

Itr2–GFPp

Fusion between Itr2 protein and green fluorescence protein

UASINO

Inositol upstream activating sequence

Rsp5p

E3 ubiquitin ligase that conjugates ubiquitin to proteins to target them for degradation

SC

Synthetic complete medium

Notes

Acknowledgements

We thank Dr. André Bastos for help in the endocytosis assays, Dr. Filipe Vilas-Boas for helpful discussions and critical reading of the manuscript and Dr. Helena Soares for help in the preparation of the figures. This work was supported by Fundação para a Ciência e a Tecnologia (FCT), Grant PEst-OE/QUI/UI0612/2013.

Compliance with ethical standards

Conflict of interest

The authors declare that there is no conflict of interest.

References

  1. Ashizawa N, Yoshida M, Aotsuka T (2000) An enzymatic assay for myo-inositol in tissue samples. J Biochem Biophys Methods 44:89–94CrossRefGoogle Scholar
  2. Becuwe M, Leon S (2014) Integrated control of transporter endocytosis and recycling by the arrestin-related protein Rod1 and the ubiquitin ligase Rsp5. Elife 3:e03307CrossRefGoogle Scholar
  3. Berridge MJ, Irvine RF (1989) Inositol phosphates and cell signalling. Nature 341:197–205CrossRefGoogle Scholar
  4. Branco MR, Marinho HS, Cyrne L, Antunes F (2004) Decrease of H2O2 plasma membrane permeability during adaptation to H2O2 in Saccharomyces cerevisiae. J Biol Chem 279:6501–6506CrossRefGoogle Scholar
  5. Camelo C, Vilas-Boas F, Cepeda AP, Real C, Barros-Martins J, Pinto F, Soares H, Marinho HS, Cyrne L (2017) Opi1p translocation to the nucleus is regulated by hydrogen peroxide in Saccharomyces cerevisiae. Yeast 34:383–395CrossRefGoogle Scholar
  6. Carman GM, Han GS (2011) Regulation of phospholipid synthesis in the yeast Saccharomyces cerevisiae. Annu Rev Biochem 80:859–883CrossRefGoogle Scholar
  7. Carman GM, Henry SA (1999) Phospholipid biosynthesis in the yeast Saccharomyces cerevisiae and interrelationship with other metabolic processes. Prog Lipid Res 38:361–399CrossRefGoogle Scholar
  8. Carman GM, Henry SA (2007) Phosphatidic acid plays a central role in the transcriptional regulation of glycerophospholipid synthesis in Saccharomyces cerevisiae. J Biol Chem 282:37293–37297CrossRefGoogle Scholar
  9. Chen M, Hancock LC, Lopes JM (2007) Transcriptional regulation of yeast phospholipid biosynthetic genes. Biochim Biophys Acta 1771:310–321CrossRefGoogle Scholar
  10. Cowart LA, Obeid LM (2007) Yeast sphingolipids: recent developments in understanding biosynthesis, regulation, and function. Biochim Biophys Acta 1771:421–431CrossRefGoogle Scholar
  11. Culbertson MR, Donahue TF, Henry SA (1976) Control of inositol biosynthesis in Saccharomyces cerevisiae: properties of a repressible enzyme system in extracts of wild-type (Ino+) cells. J Bacteriol 126:232–242PubMedPubMedCentralGoogle Scholar
  12. Dulic V, Egerton M, Elguindi I, Raths S, Singer B, Riezman H (1991) Yeast endocytosis assays. Methods Enzymol 194:697–710CrossRefGoogle Scholar
  13. Folmer V, Pedroso N, Matias AC, Lopes SCDN, Antunes F, Cyrne L, Marinho HS (2008) H2O2 induces rapid biophysical and permeability changes in the plasma membrane of Saccharomyces cerevisiae. Biochim Biophys Acta 1778:1141–1147CrossRefGoogle Scholar
  14. Gaspar ML, Aregullin MA, Jesch SA, Henry SA (2006) Inositol induces a profound alteration in the pattern and rate of synthesis and turnover of membrane lipids in Saccharomyces cerevisiae. J Biol Chem 281:22773–22785CrossRefGoogle Scholar
  15. Greenberg ML, Lopes JM (1996) Genetic regulation of phospholipid biosynthesis in Saccharomyces cerevisiae. Microbiol Rev 60:1–20PubMedPubMedCentralGoogle Scholar
  16. Grossmann G, Malinsky J, Stahlschmidt W, Loibl M, Weig-Meckl I, Frommer WB, Opekarova M, Tanner W (2008) Plasma membrane microdomains regulate turnover of transport proteins in yeast. J Cell Biol 183:1075–1088CrossRefGoogle Scholar
  17. Henry SA, Kohlwein SD, Carman GM (2012) Metabolism and regulation of glycerolipids in the yeast Saccharomyces cerevisiae. Genetics 190:317–349CrossRefGoogle Scholar
  18. Hitchcock AL, Auld K, Gygi SP, Silver PA (2003) A subset of membrane-associated proteins is ubiquitinated in response to mutations in the endoplasmic reticulum degradation machinery. Proc Natl Acad Sci USA 100:12735–12740CrossRefGoogle Scholar
  19. Huh WK, Falvo JV, Gerke LC, Carroll AS, Howson RW, Weissman JS, O’Shea EK (2003) Global analysis of protein localization in budding yeast. Nature 425:686–691CrossRefGoogle Scholar
  20. Jesch SA, Zhao X, Wells MT, Henry SA (2005) Genome-wide analysis reveals inositol, not choline, as the major effector of Ino2p–Ino4p and unfolded protein response target gene expression in yeast. J Biol Chem 280:9106–9118CrossRefGoogle Scholar
  21. Kelley MJ, Bailis AM, Henry SA, Carman GM (1988) Regulation of phospholipid biosynthesis in Saccharomyces cerevisiae by inositol. Inositol is an inhibitor of phosphatidylserine synthase activity. J Biol Chem 263:18078–18085PubMedGoogle Scholar
  22. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685CrossRefGoogle Scholar
  23. Lai K, McGraw P (1994) Dual control of inositol transport in Saccharomyces cerevisiae by irreversible inactivation of permease and regulation of permease synthesis by INO2, INO4, and OPI1. J Biol Chem 269:2245–2251PubMedGoogle Scholar
  24. Lai K, Bolognese CP, Swift S, McGraw P (1995) Regulation of inositol transport in Saccharomyces cerevisiae involves inositol-induced changes in permease stability and endocytic degradation in the vacuole. J Biol Chem 270:2525–2534CrossRefGoogle Scholar
  25. Lin CH, MacGurn JA, Chu T, Stefan CJ, Emr SD (2008) Arrestin-related ubiquitin-ligase adaptors regulate endocytosis and protein turnover at the cell surface. Cell 135:714–725CrossRefGoogle Scholar
  26. Loewen CJ, Gaspar ML, Jesch SA, Delon C, Ktistakis NT, Henry SA, Levine TP (2004) Phospholipid metabolism regulated by a transcription factor sensing phosphatidic acid. Science 304:1644–1647CrossRefGoogle Scholar
  27. Marinho HS, Cyrne L, Cadenas E, Antunes F (2013) H2O2 delivery to cells: steady-state versus bolus addition. Methods Enzymol 526:159–173CrossRefGoogle Scholar
  28. Marinho HS, Real C, Cyrne L, Soares H, Antunes F (2014) Hydrogen peroxide sensing, signaling and regulation of transcription factors. Redox Biol 2:535–562CrossRefGoogle Scholar
  29. Matias AC, Pedroso N, Teodoro N, Marinho HS, Antunes F, Nogueira JM, Herrero E, Cyrne L (2007) Down-regulation of fatty acid synthase increases the resistance of Saccharomyces cerevisiae cells to H2O2. Free Radic Biol Med 43:1458–1465CrossRefGoogle Scholar
  30. Miyashita M, Shugyo M, Nikawa J (2003) Mutational analysis and localization of the inositol transporters of Saccharomyces cerevisiae. J Biosci Bioeng 96:291–297CrossRefGoogle Scholar
  31. Nikawa J, Tsukagoshi Y, Yamashita S (1991) Isolation and characterization of two distinct myo-inositol transporter genes of Saccharomyces cerevisiae. J Biol Chem 266:11184–11191PubMedGoogle Scholar
  32. Nikawa J, Hosaka K, Yamashita S (1993) Differential regulation of two myo-inositol transporter genes of Saccharomyces cerevisiae. Mol Microbiol 10:955–961CrossRefGoogle Scholar
  33. Nikko E, Pelham HR (2009) Arrestin-mediated endocytosis of yeast plasma membrane transporters. Traffic 10:1856–1867CrossRefGoogle Scholar
  34. Oliveira-Marques V, Silva T, Cunha F, Covas G, Marinho HS, Antunes F, Cyrne L (2013) A quantitative study of the cell-type specific modulation of c-Rel by hydrogen peroxide and TNF-alpha. Redox Biol 1:347–352CrossRefGoogle Scholar
  35. Paulick MG, Bertozzi CR (2008) The glycosylphosphatidylinositol anchor: a complex membrane-anchoring structure for proteins. Biochemistry 47:6991–7000CrossRefGoogle Scholar
  36. Pedroso N, Matias AC, Cyrne L, Antunes F, Borges C, Malhó R, de Almeida RF, Herrero E, Marinho HS (2009) Modulation of plasma membrane lipid profile and microdomains by H2O2 in Saccharomyces cerevisiae. Free Radic Biol Med 46:289–298CrossRefGoogle Scholar
  37. Pedroso N, Gomes-Alves P, Marinho HS, Brito VB, Boada C, Antunes F, Herrero E, Penque D, Cyrne L (2012) The plasma membrane-enriched fraction proteome response during adaptation to hydrogen peroxide in Saccharomyces cerevisiae. Free Radic Res 46:1267–1279CrossRefGoogle Scholar
  38. Peterson GL (1977) A simplification of the protein assay method of Lowry et al. which is more generally applicable. Anal Biochem 83:346–356CrossRefGoogle Scholar
  39. Polo S, Di Fiore PP (2008) Finding the right partner: science or ART? Cell 135:590–592CrossRefGoogle Scholar
  40. Robinson KS, Lai K, Cannon TA, McGraw P (1996) Inositol transport in Saccharomyces cerevisiae is regulated by transcriptional and degradative endocytic mechanisms during the growth cycle that are distinct from inositol-induced regulation. Mol Biol Cell 7:81–89CrossRefGoogle Scholar
  41. Schindelin J, Arganda-Carreras I, Frise E, Kaynig V, Longair M, Pietzsch T, Preibisch S, Rueden C, Saalfeld S, Schmid B, Tinevez JY, White DJ, Hartenstein V, Eliceiri K, Tomancak P, Cardona A (2012) Fiji: an open-source platform for biological-image analysis. Nat Methods 9:676–682CrossRefGoogle Scholar
  42. Schneider S (2015) Inositol transport proteins. FEBS Lett 589:1049–1058CrossRefGoogle Scholar
  43. Sousa-Lopes A, Antunes F, Cyrne L, Marinho HS (2004) Decreased cellular permeability to H2O2 protects Saccharomyces cerevisiae cells in stationary phase against oxidative stress. FEBS Lett 578:152–156CrossRefGoogle Scholar
  44. Strahl T, Thorner J (2007) Synthesis and function of membrane phosphoinositides in budding yeast Saccharomyces cerevisiae. Biochim Biophys Acta 1771:353–404CrossRefGoogle Scholar
  45. Tsui MM, York JD (2010) Roles of inositol phosphates and inositol pyrophosphates in development, cell signaling and nuclear processes. Adv Enzyme Regul 50:324–337CrossRefGoogle Scholar
  46. Wiederkehr A, Avaro S, Prescianotto-Baschong C, Haguenauer-Tsapis R, Riezman H (2000) The F-box protein Rcy1p is involved in endocytic membrane traffic and recycling out of an early endosome in Saccharomyces cerevisiae. J Cell Biol 149:397–410CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Centro de Química e Bioquímica, Faculdade de CiênciasUniversidade de LisboaLisbonPortugal
  2. 2.Departamento de Química e Bioquímica, Faculdade de CiênciasUniversidade de LisboaLisbonPortugal

Personalised recommendations