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The acid phosphatase Pho5 of Saccharomyces cerevisiae is not involved in polyphosphate breakdown

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Abstract

Inorganic polyphosphate is involved in architecture and functioning of yeast cell wall. The strain of Saccharomyces cerevisiae constitutively overexpressing acid phosphatase Pho5 was constructed for studying the Pho5 properties and its possible participation in polyphosphate metabolism. The parent strain was transformed by the vector carrying the PHO5 gene under a strong constitutive promoter of glyceraldehyde-3-phosphate dehydrogenase of S. cerevisiae. The culture liquid and biomass of transformant strain contained approximately equal total acid phosphatase activity. The levels of acid phosphatase activity associated with the cell wall and culture liquid increased in the transformant strain compared to the parent strain ~ 10- and 20-fold, respectively. The Pho5 preparation (specific activity of 46 U/mg protein and yield of 95 U/L) was obtained from culture liquid of overproducing strain. The overproducing strain had no changes in polyphosphate level. The activity of Pho5 with long-chained polyP was negligible. We concluded that Pho5 is not involved in polyphosphate metabolism. Purified Pho5 showed a similar activity with p-nitrophenylphosphate, ATP, ADP, glycerophosphate, and glucose-6-phosphate. The substrate specificity of Pho5 and its extracellular localization suggest its function: the hydrolysis of organic compounds with phosphoester bonds at phosphate limitation.

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References

  • Albi T, Serrano A (2016) Inorganic polyphosphate in the microbial world. Emerging roles for a multifaceted biopolymer. World J Microbiol Biotechnol 32:27. https://doi.org/10.1007/s11274-015-1983-2

    Article  CAS  PubMed  Google Scholar 

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

    Article  CAS  Google Scholar 

  • Anemaet IG, van Heusden GP (2014) Transcriptional response of Saccharomyces cerevisiae to potassium starvation. BMC Genomics 15:1040. https://doi.org/10.1186/1471-2164-15-1040

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Bergmeyer HU, Gawehn K, Grassl M (1974) Enzymes as biochemical reagent. In: Bergmeyer HU (ed) Method of enzymatic analysis, vol I, 2nd edn. Academic Press, Inc., New York, NY, pp 495–496

    Google Scholar 

  • Clotet J (2017) Polyphosphate: popping up from oblivion. Curr Genet 63:15–18

    Article  Google Scholar 

  • Eldarov MA, Baranov MV, Dumina MV, Shgun AA, Andreeva NA, Trilisenko LV, Kulakovskaya TV, Ryasanova LP, Kulaev IS (2013) Polyphosphates and exopolyphosphatase activities in the yeast Saccharomyces cerevisiae under overexpression of homologous and heterologous PPN1 genes. Biochem Mosc 78:946–953

    Article  CAS  Google Scholar 

  • Eskes E, Deprez MA, Wilms T, Winderickx J (2018) pH homeostasis in yeast; the phosphate perspective. Curr Genet 64:155–161

    Article  CAS  Google Scholar 

  • Gietz RD, Schiest RH (2007) High-efficiency yeast transformation using the LiAc/SS carrier DNA/PEG method. Nat Protoc 2:31–34

    Article  CAS  Google Scholar 

  • Halonen P, Baykov AA, Goldman A, Lahti R, Cooperman BS (2002) Single-turnover kinetics of Saccharomyces cerevisiae inorganic pyrophosphatase. Biochemistry 41:12025–12031

    Article  CAS  Google Scholar 

  • Kalebina TS, Egorov SN, Arbatskii NP, Bezsonov EE, Gorkovskii AA, Kulaev IS (2008) The role of high-molecular-weight polyphosphates in activation of glucan transferase Bgl2p from Saccharomyces cerevisiae cell wall. Dokl Biochem Biophys 420:142–145

    Article  CAS  Google Scholar 

  • Kane PM (2016) Proton transport and pH control in fungi. Adv Exp Med Biol 892:33–68

    Article  CAS  Google Scholar 

  • Kennedy EJ, Pillus L, Ghosh G (2005) Pho5p and newly identified nucleotide pyrophosphatases/phosphodiesterases regulate extracellular nucleotide phosphate metabolism in Saccharomyces cerevisiae. Eukaryot Cell 4:1892–1901

    Article  CAS  Google Scholar 

  • Kizawa K, Aono T, Ohtomo R (2017) PHO8 gene coding alkaline phosphatase of Saccharomyces cerevisiae is involved in polyphosphate metabolism. J Gen Appl Microbiol 62:297–302

    Article  Google Scholar 

  • Korber P, Barbaric S (2014) The yeast PHO5 promoter: from single locus to systems biology of a paradigm for gene regulation through chromatin. Nucl Acids Res 42:10888–10902

    Article  CAS  Google Scholar 

  • Kornberg RD (2005) Mediator and the mechanism of transcriptional activation. Trends Biochem Sci 30:235–239

    Article  CAS  Google Scholar 

  • Kozulić B, Barbarić S, Ries B, Mildner P (1984) Study of the carbohydrate part of yeast acid phosphatase. Biochem Biophys Res Commun 122:1083–1090

    Article  Google Scholar 

  • Kulaev IS, Vagabov VM, Kulakovskaya TV (2004) The Biochemistry of Inorganic Polyphosphates. John Wiley & Sons Ltd, Chichester

    Book  Google Scholar 

  • Kumar A, Gangaiah D, Torrelles JB, Rajashekara G (2016) Polyphosphate and associated enzymes as global regulators of stress response and virulence in Campylobacter jejuni. World J Gastroenterol 22:7402–7414

    Article  CAS  Google Scholar 

  • Magnelli PE, Bielik AM, Guthrie EP (2011) Identification and characterization of protein glycosylation using specific endo- and exoglycosidases. J Vis Exp 58:3749. https://doi.org/10.3791/3749

    Article  CAS  Google Scholar 

  • Nosaka K (1990) High affinity of acid phosphatase encoded by PHO3 gene in Saccharomyces cerevisiae for thiamin phosphates. Biochem Biophys Acta 1037:147–154

    CAS  PubMed  Google Scholar 

  • Oshima Y (1997) The phosphatase system in Saccharomyces cerevisiae. Gen Genet Syst 72:323–334

    Article  CAS  Google Scholar 

  • Pondugula S, Neef DW, Voth WP, Darst RP, Dhasarathy A, Reynolds MM, Takahata S, Stillman DJ, Kladde MP (2009) Coupling phosphate homeostasis to cell cycle-specific transcription: mitotic activation of Saccharomyces cerevisiae PHO5 by Mcm1 and forkhead proteins. Mol Cell Biol 29:4891–4905

    Article  CAS  Google Scholar 

  • Ramos CL, Gomes FM, Girard-Dias W, Almeida FP, Albuquerque PC, Kretschmer M, Kronstad JW, Frases S, de Souza W, Rodrigues ML, Miranda K (2017) Phosphorus-rich structures and capsular architecture in Cryptococcus neoformans. Future Microbiol 12:227–238

    Article  CAS  Google Scholar 

  • Rao NN, Gómez-García MR, Kornberg A (2009) Inorganic polyphosphate: essential for growth and survival. Annu Rev Biochem 78:605–647

    Article  CAS  Google Scholar 

  • Rubin GM (1973) The nucleotide sequence of Saccharomyces cerevisiae 5.8 S ribosomal ribonucleic acid. J Biol Chem 11:3860–3875

    Google Scholar 

  • Ryazanova L, Zvonarev A, Rusakova T, Dmitriev V, Kulakovskaya T (2016) Manganese tolerance in yeasts involves polyphosphate, magnesium, and vacuolar alterations. Folia Microbiol (Praha) 61:311–317

    Article  CAS  Google Scholar 

  • Secco D, Wang C, Shou H, Whelan J (2012) Phosphate homeostasis in the yeast Saccharomyces cerevisiae, the key role of the SPX domain-containing proteins. FEBS Lett 586:289–295

    Article  CAS  Google Scholar 

  • Shabalin YA, Kulaev IS (1989) Solubilization and properties of yeast dolichylpyrophosphate:polyphosphate phosphotransferase. Biokhimia (Moscow) 54:68–75

    CAS  Google Scholar 

  • Shnyreva MG, Petrova EV, Egorov SN, Hinnen A (1996) Biochemical properties and excretion behavior of repressible acid phosphatases with altered subunit composition. Microbiol Res 151:291–300

    Article  CAS  Google Scholar 

  • Sikorski RS, Hieter P (1989) A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae. Genetic 122:19–27

    CAS  Google Scholar 

  • Skelton TP, Zeng C, Nocks A, Stamenkovic I (1998) Glycosylation provides both stimulatory and inhibitory effects on cell surface and soluble CD44 binding to hyaluronan. J Cell Biol 140:431–446

    Article  CAS  Google Scholar 

  • Spencer F, Ketner G, Connelly C, Hieter P (1993) Targeted recombination-based cloning and manipulation of large DNA segments in yeast. Methods 5:161–175

    Article  CAS  Google Scholar 

  • Toh-e A, Ueda Y, Kakimoto S-I, Oshima Y (1973) Isolation and characterization of acid phosphatase mutants in Saccharomyces cerevisiae. J Bacteriol 113:727–738

    CAS  PubMed Central  Google Scholar 

  • Vagabov VM, Trilisenko LV, Kulaev IS (2000) Dependence of inorganic polyphosphate chain length on the orthophosphate content in the culture medium of the yeast Saccharomyces cerevisiae. Biochem Mosc 65:349–354

    CAS  Google Scholar 

  • Yadav KK, Singh N, Rajasekharan R (2016) Responses to phosphate deprivation in yeast cells. Curr Genet 62(2):301–307. https://doi.org/10.1007/s00294-015-0544-4

    Article  CAS  PubMed  Google Scholar 

  • Zvonarev AN, Crowley DE, Ryazanova LP, Lichko LP, Rusakova TG, Kulakovskaya TV, Dmitriev VV (2017) Cell wall canals formed upon growth of Candida maltosa in the presence of hexadecane are associated with polyphosphates. FEMS Yeast Res 17. https://doi.org/10.1093/femsyr/fox026

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Acknowledgements

The authors thank Elena Makeeva for her help with preparing the manuscript.

Funding

This study was supported in part by the Russian Foundation for Basic Research (grant no. 17-04-00822).

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Authors and Affiliations

  1. FRC Pushchino Center for Biological Research, Skryabin Institute of Biochemistry and Physiology of Microorganisms, Russian Academy of Sciences, pr. Nauki 5, Pushchino, Moscow region, 142290, Russia

    Nadeshda Andreeva, Larisa Ledova, Lubov Ryasanova & Tatiana Kulakovskaya

  2. Institute of Bioengineering, Research Center of Biotechnology of the Russian Academy of Sciences, Leninsky prosp. 33-2, Moscow, 119071, Russia

    Michail Eldarov

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  1. Nadeshda Andreeva
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  2. Larisa Ledova
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  3. Lubov Ryasanova
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  4. Tatiana Kulakovskaya
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  5. Michail Eldarov
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Correspondence to Tatiana Kulakovskaya.

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Andreeva, N., Ledova, L., Ryasanova, L. et al. The acid phosphatase Pho5 of Saccharomyces cerevisiae is not involved in polyphosphate breakdown. Folia Microbiol 64, 867–873 (2019). https://doi.org/10.1007/s12223-019-00702-6

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  • DOI: https://doi.org/10.1007/s12223-019-00702-6

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