Skip to main content
Log in

Inorganic Polyphosphates and Exopolyphosphatases in Cell Compartments of the Yeast Saccharomyces cerevisiae Under Inactivation of PPX1 and PPN1 Genes

  • Published:
Bioscience Reports

Abstract

Purified fractions of cytosol, vacuoles, nuclei, and mitochondria of Saccharomyces cerevisiae possessed inorganic polyphosphates with chain lengths characteristic of each individual compartment. The most part (80–90%) of the total polyphosphate level was found in the cytosol fractions. Inactivation of a PPX1 gene encoding ~40-kDa exopolyphosphatase substantially decreased exopolyphosphatase activities only in the cytosol and soluble mitochondrial fraction, the compartments where PPX1 activity was localized. This inactivation slightly increased the levels of polyphosphates in the cytosol and vacuoles and had no effect on polyphosphate chain lengths in all compartments. Exopolyphosphatase activities in all yeast compartments under study critically depended on the PPN1 gene encoding an endopolyphosphatase. In the single PPN1 mutant, a considerable decrease of exopolyphosphatase activity was observed in all the compartments under study. Inactivation of PPN1 decreased the polyphosphate level in the cytosol 1.4-fold and increased it 2- and 2.5-fold in mitochondria and vacuoles, respectively. This inactivation was accompanied by polyphosphate chain elongation. In nuclei, this mutation had no effect on polyphosphate level and chain length as compared with the parent strain CRY. In the double mutant of PPX1 and PPN1, no exopolyphosphatase activity was detected in the cytosol, nuclei, and mitochondria and further elongation of polyphosphates was observed in all compartments.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig 1

Similar content being viewed by others

References

  1. Kulaev IS (1979) The biochemistry of inorganic polyphosphates. Wiley, New York

    Google Scholar 

  2. Kulaev IS, Vagabov VM, Kulakovskaya TV (2004) The biochemistry of inorganic polyphosphates. Wiley, New York

    Google Scholar 

  3. Urech K, Durr M, Boller T, Wiemken A (1978) Localization of polyphosphate in vacuoles of Saccharomyces cerevisiae. Arch Microbiol 116:274–278

    Article  Google Scholar 

  4. Lichko LP, Okorokov LA, Kulaev IS (1982) Participation of vacuoles in regulation of K+, Mg2+ and orthophosphate ions in cytoplasm of the yeast Saccharomyces carlsbergensis. Arch Microbiol 132:289–293

    Article  CAS  Google Scholar 

  5. Kornberg A, Rao NN, Ault-Riché D (1999) Inorganic polyphosphate: a molecule with many functions. Ann Rev Biochem 68:89–125

    Article  PubMed  CAS  Google Scholar 

  6. Offenbacher S, Kline H (1984) Evidence for polyphosphate in phosphorylated nonhistone nuclear proteins. Arch Biochem Biophys 231:114–123

    Article  PubMed  CAS  Google Scholar 

  7. Beauvoit B, Rigonlet M, Guerin B, Canioni P (1989) Polyphosphates as a source of high energy phosphates in yeast mitochondria: a P-NMR study. FEBS Lett 5252:17–22

    Article  Google Scholar 

  8. Reush RN (1992) Biological complexes of poly-β-hydroxybutyrate. FEMS Rev 103:119–130

    Google Scholar 

  9. Lichko LP, Andreeva NA, Kulakovskaya TV, Kulaev IS (2003) Exopolyphosphatases of the yeast Saccharomyces cerevisiae. FEMS Yeast Res 3:233–238

    Article  PubMed  CAS  Google Scholar 

  10. Wurst H, Shiba T, Kornberg A (1995) The gene for a major exopolyphosphatase of Saccharomyces cerevisiae. J Bacteriol 177:898–906

    PubMed  CAS  Google Scholar 

  11. Kumble KD, Kornberg A (1996) Endopolyphosphatases for long chain polyphosphate in yeast and mammals. J Biol Chem 271:27146–27151

    Article  PubMed  CAS  Google Scholar 

  12. Sethuraman A, Rao NN, Kornberg A (2001) The endopolyphosphatase gene: essential in Saccharomyces cerevisiae. Proc Natl Acad Sci USA 98:8542–8547

    Article  PubMed  CAS  Google Scholar 

  13. Ogawa N, DeRisi J, Brown PO (2000) New components of a system for phosphate accumulation and polyphosphate metabolism in Saccharomyces cerevisiae revealed by genomic expression analysis. Mol Biol Cell 11:4309–4321

    PubMed  CAS  Google Scholar 

  14. Lichko LP, Kulakovskaya TV, Kulaev IS (2004) Inactivation of endopolyphosphatase gene PPN1 results in inhibition of expression of exopolyphosphatase PPX1 and high-molecular-mass exopolyphosphatase not encoded by PPX1 in Saccharomyces cerevisiae. Biochim Biophys Acta 1674:98–102

    PubMed  CAS  Google Scholar 

  15. Shi X, Kornberg A (2005) Endopolyphosphatase in Saccharomyces cerevisiae undergoes posttranslational activations to produce shortchain polyphosphates. FEBS Lett 579:2014–2018

    Article  PubMed  CAS  Google Scholar 

  16. Andreea NA, Okorokov LA (1993) Purification and characterization of highly active and stable polyphosphatase from Saccharomyces cerevisiae cell envelope. Yeast 9:127–139

    Article  PubMed  CAS  Google Scholar 

  17. Wurst H, Kornberg A (1994) A soluble exopolyphosphatase of Saccharomyces cerevisiae. J Biol Chem 269:10996–11001

    PubMed  CAS  Google Scholar 

  18. Andreeva N, Kulakovskaya T, Sidorov I, Karpov A, Kulaev I (1998) Purification and properties of polyphosphatase from Saccharomyces cerevisiae cytosol. Yeast 14:383–390

    Article  PubMed  CAS  Google Scholar 

  19. Lichko LP, Kulakovskaya TV, Kulaev IS (2003) Nuclear exopolyphosphatase of Saccharomyces cerevisiae is not encoded by PPX1 gene encoding the major yeast exopolyphosphatase. FEMS Yeast Res 3:113–117

    PubMed  CAS  Google Scholar 

  20. Lichko LP, Kulakovskaya TV, Dmitriev VV, Kulaev IS (1995) Saccharomyces cerevisiae nuclei possess polyphosphatase activity. Biochemistry (Moscow) 60:1465–1468

    Google Scholar 

  21. Lichko LP, Okorokov LA (1984) Some properties of membrane-bound, solubilized and reconstituted into liposomes H+-ATPase of vacuoles of Saccharomyces carlsbergensis. FEBS Lett 174:233–237

    Article  PubMed  CAS  Google Scholar 

  22. Lichko LP, Kulakovskaya TV, Kulaev IS (1998) Membrane-bound and soluble polyphosphatases of mitochondria of Saccharomycres cerevisiae: identification and comparative characterization. Biochim Biophys Acta 1372:153–162

    Article  PubMed  CAS  Google Scholar 

  23. Guerin B, Lable P, Somlo M (1981) Preparation of yeast mitochondria (Saccharomyces cerevisiae) with good P/O and respiratory control ratios. Methods Enzymol 55:149–159

    Article  Google Scholar 

  24. Kotelnikova AV, Zvjagilskaya RA (1973) The biochemistry of yeast mitochondria. Science, Moscow

  25. Mejer MMC, Boonstra J, Verkleij AJ, Verrips CT (1998) Glucose repression in Saccharomyces cerevisiae is related to the glucose concentration rather than glucose flux. J Biol Chem 278:24102–24107

    Article  Google Scholar 

  26. Andreeva NA, Kulakovskaya TV, Kulaev IS (1996) Purification and characterization of polyphosphatase from Saccharomyces cerevisiae cytosol. Biochemistry (Moscow) 61:1213–1220

    Google Scholar 

  27. Kulaev IS, Belozersky AN, Kritsky MS, Kokurina NA (1960) Polyphosphates of the fruiting bodies of the mushroom and false morel. Dokl Akad Nauk SSSR 130:667–670 (in Russian)

    CAS  Google Scholar 

  28. Kumble KD, Kornberg A (1995) Inorganic polyphosphate in mammalian cells and tissues. J Biol Chem 270:5818–5822

    Article  PubMed  CAS  Google Scholar 

  29. Bensadoun A, Weinstein D (1976) Assay of proteins in the presence of interfering materials. Anal Biochem 70:241–250

    Article  PubMed  CAS  Google Scholar 

  30. Lichko LP, Pestov NA, Kulakovskaya TV, Kulaev IS (2003) Effect of PPX1 inactivation on the exopolyphosphatase spectra in cytosol and mitochondria of the yeast. Biochemistry (Moscow) 68:740–746

    Article  CAS  Google Scholar 

  31. Trilisenko LV, Vagabov VM, Kulaev IS (2002) The content and chain length of polyphosphate from vacuoles of Saccharomyces cerevisiae VKM Y-1173. Biochemistry (Moscow) 67:592–595

    Article  CAS  Google Scholar 

  32. Wiemken A, Schellenberg M, Urech R (1979) Vacuoles: the sole compartments of digestive enzymes in yeast (Saccharomyces cerevisiae). Arch Microbiol 123:23–25

    Article  CAS  Google Scholar 

  33. Kulaev IS, Krasheninnikov IA, Kokurina NA (1966) The localization of inorganic polyphosphates and nucleotides in mycelia of Neurospora crassa. Biokhimiya 31:850–858 (in Russian)

    CAS  Google Scholar 

  34. Matile P (1978) Biochemistry and function of vacuoles. Ann Rev Plant Physiol 29:193–213

    Article  CAS  Google Scholar 

  35. Nunez CG, Callieri AS (1989) Studies on the polyphosphate cycle in Candida utilis. Effect of dilution rate and nitrogen source in continuous culture. Appl Microbiol Biotechnol 31:562–566

    Article  CAS  Google Scholar 

  36. Lichko LP, Okorokov LA, Kulaev IS (1982) Participation of vacuoles in regulation of K+, Mg2+ and orthophosphate ions in cytoplasm of the yeast Saccharomyces carlsbergensis. Arch Microbiol 132:289–293

    Article  CAS  Google Scholar 

  37. Vorisek J, Knotkova A, Kotyk A (1982) Fine cytochemical localization of polyphosphates in the yeast Saccharomyces cerevisiae. ZBL Microbiol 137:421–432

    CAS  Google Scholar 

  38. Guranowski A, Starzynska E, Barnes LD, Robinson AK, Liu S (1998) Adenosine 5'-tetraphosphate phosphohydrolase activity is an inherent property of soluble exopolyphosphatase from Saccharomyces cerevisiae. Biochim Biophys Acta 1380:232–238

    PubMed  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported by the Russian Foundation for Basic Research (Grant 05-04-48175) and a grant supporting the leading scientific schools 1382.2003.4. We appreciate the valuable technical assistance of L. Mihailina and N. Kosenkova.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Lidiya Lichko.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Lichko, L., Kulakovskaya, T., Pestov, N. et al. Inorganic Polyphosphates and Exopolyphosphatases in Cell Compartments of the Yeast Saccharomyces cerevisiae Under Inactivation of PPX1 and PPN1 Genes. Biosci Rep 26, 45–54 (2006). https://doi.org/10.1007/s10540-006-9003-2

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10540-006-9003-2

Keywords

Navigation