AiM  Vol.4 No.8 , June 2014
Analysis of Volutin Granule Formation in Saccharomyces cerevisiae
ABSTRACT

The budding yeast Saccharomyces cerevisiae serves as an effective model organism for many cellular pathways including phosphate transport, accumulation, and storage. In S. cerevisiae, phosphate is actively transported across the plasma membrane via several phosphate carriers and is then transported into the acidic vacuole (roughly equivalent to the mammalian lysosome with degradative functions but with additional storage functions, such as calcium) where it is synthesized into volutin, a storage form of polyphosphate, found in many organisms. We have been studying volutin granule formation in wild type cells to determine the physiological requirements for formation and in mutants to determine the pathway by which the volutin biosynthetic proteins are transported to the vacuole. Undertaking an analysis of volutin formation in yeast vacuoles by blocking vacuole function with pharmacological agents, such as ionomycin and CCCP, we see that vacuole pH as well as vacuolar calcium seems critical for volutin formation. Different blocks in vacuolar protein sorting have differential effects on volutin granule accumulation, with volutin granule formation seen in all mutant strains thus far tested, except for vps33, a mutant cell strain lacking all vacuolar structure. Our data are consistent with pleiotrophic effects of vacuolar physiological function blocks leading to a decrease in volutin formation.


Cite this paper
Marshall, P. , Rosa, D. , Sanchez, L. and Starr, M. (2014) Analysis of Volutin Granule Formation in Saccharomyces cerevisiae. Advances in Microbiology, 4, 465-473. doi: 10.4236/aim.2014.48051.
References
[1]   Lenburg, M.E. and O’Shea, E.K. (1996) Signaling Phosphate Starvation. Trends in Biochemical Sciences, 21, 383-387. http://dx.doi.org/10.1016/0968-0004(96)10048-7

[2]   Urech, K., Durr, M., Boller, T., Wiemken, A. and Schwencke, J. (1978) Localization of Polyphosphate in Vacuoles of Saccharomyces cerevisiae. Archives of Microbiology, 116, 275-278.
http://dx.doi.org/10.1007/BF00417851

[3]   Saito, K., Ohtomo, R., Kuga-Uetake, Y., Aono, T. and Saito, M. (2005) Direct Labeling of Polyphosphate at the Ultrastructural Level in Saccharomyces cerevisiae by Using The Affinity of the Polyphosphate Binding Domain of Escherichia Coli Exopolyphosphatase. Applied and Environmental Microbiology, 71, 5692-5701.

[4]   Ogawa, N., DeRisi, J. and Brown, P.O. (2000) New Components of a System for Phosphate Accumulation and Polyphosphate Metabolism in Saccharomyces cerevisiae Revealed by Genomic Expression Analysis. Molecular Biology of the Cell, 11, 4309-4321.
http://dx.doi.org/10.1091/mbc.11.12.4309

[5]   Freimoser, F.M., Hurlimann, H.C., Jakob, C.A., Werner, T.P. and Amrhein, N. (2006) Systematic Screening of Polyphosphate(poly P) Levels in Yeast Mutant Cells Reveals Strong Interdependence with Primary Metabolism. Genome Biology, 7, R109.

[6]   Docampo, R., de Souza, W., Miranda, K., Rohloff, P. and Moreno, S.N. (2005) Acidocalcisomes—Conserved from Bacteria to Man. Nature Reviews Microbiology, 3, 251-261.

[7]   Persson, B.L., Lagerstedt, J.O., Pratt, J.R., Pattison-Granberg, J., Lundh, K., Shokrollahzadeh, S. and Lundh, F. (2003) Regulation of Phosphate Acquisition in Saccharomyces cerevisiae. Current Genetics, 43, 225-244. http://dx.doi.org/10.1007/s00294-003-0400-9

[8]   Secco, D., Wang, C., Shou, H. and Whelan, J. (2012) Phosphate Homeostasis in the Yeast Saccharomyces cerevisiae, the Key Role of the SPX Domain-Containing Proteins. FEBS Letters, 586, 289-295. http://dx.doi.org/10.1016/j.febslet.2012.01.036

[9]   Secco, D., Wang, C., Arpat, B.A., Wang, Z., Poirier, Y., Tyerman, S.D., Wu, P., Shou, H. and Whelan, J. (2012) The Emerging Importance of the SPX Domain-Containing Proteins in Phosphate Homeostasis. New Phytologist, 193, 842-851. http://dx.doi.org/10.1111/j.1469-8137.2011.04002.x

[10]   Uttenweiler, A., Schwarz, H., Neumann, H. and Mayer, A. (2007) The Vacuolar Transporter Chaperone (VTC) Complex Is Required for Microautophagy. Molecular Biology of the Cell, 18, 166-175.

[11]   Hothorn, M., Neumann, H., Lenherr, E.D., Wehner, M., Rybin, V., Hassa, P.O., Uttenweiler, A., Reinhardt, M., Schmidt, A., Seiler, J., Ladurner, A.G., Herrmann, C., Scheffzek, K. and Mayer, A. (2009) Catalytic Core of a Membrane-Associated Eukaryotic Polyphosphate Polymerase. Science, 324, 513-516. http://dx.doi.org/10.1126/science.1168120

[12]   Vagabov, V.M., Trilisenko, L.V., Shchipanova, I.N., Sibel’dina, L.A. and Kulaev, I.S. (1998) Change in Inorganic Polyphosphate Chain Length Depending on the Stage of Saccharomyces cerevisiae Growth. Mikrobiologiia, 67, 188-193.

[13]   Vagabov, V.M., Trilisenko, L.V. and Kulaev, I.S. (2000) Dependence of Inorganic Polyphosphate Chain Length on the Orthophosphate Content in the Culture Medium of the Yeast Saccharomyces cerevisiae. Biochemistry (Mosc), 65, 349-354.

[14]   Trilisenko, L.V., Andreeva, N.A., Kulakovskaya, T.V., Vagabov, V.M. and Kulaev, I.S. (2003) Effect of Inhibitors on Polyphosphate Metabolism in the Yeast Saccharomyces cerevisiae under Hypercompensation Conditions. Biochemistry (Mosc), 68, 577-581.

[15]   Vagabov, V.M., Trilisenko, L.V., Kulakovskaya, T.V. and Kulaev, I.S. (2008) Effect of a Carbon Source on Polyphosphate Accumulation in Saccharomyces cerevisiae. FEMS Yeast Research, 8, 877-882. http://dx.doi.org/10.1111/j.1567-1364.2008.00420.x

[16]   Trilisenko, L., Tomashevsky, A., Kulakovskaya, T. and Kulaev, I. (2013) V-ATPase Dysfunction Suppresses Polyphosphate Synthesis in Saccharomyces cerevisiae. Folia Microbiologica (Praha), 58, 437-441. http://dx.doi.org/10.1007/s12223-013-0226-x

[17]   Trilisenko, L.V., Kochetkova, O.Y., Vagabov, V.M. and Kulaev, I.S. (2010) Effect of Cycloheximide, Iodoacetamide, and Antimycin A on Inorganic Phosphate Synthesis in Saccharomyces cerevisiae VKM Y-1173. Microbiology, 79, 23-29. http://dx.doi.org/10.1134/S0026261710010030

[18]   Vagabov, V.M., Trilisenko, L.V., Kochetkova, O.Y., Ilchenko, A.P. and Kulaev, I.S. (2011) Effect of m-Carbonyl Cyanide 3-Chlorophenylhydrazone on Inorganic Polyphosphates Synthesis in Saccharomy
ces cerevisiae under Different Growth Conditions. Microbiology, 80, 15-20.
http://dx.doi.org/10.1134/S0026261711010176

[19]   Breus, N.A., Riazanova, L.P., Suzina, N.E., Kulakovskaia, N.V., Valiakhmetov, A.I., Iashin, V.A., Sorokin, V.V. and Kulaev, I.S. (2011) Accumulation of Inorganic Polyphosphates in Saccharomyces cerevisiae under Nitrogen Deprivation: Stimulation by Magnesium Ions and Peculiarities of Localization. Mikrobiologiia, 80, 624-630. http://dx.doi.org/10.1134/S002626171105002X

[20]   Rothman, J.H. and Stevens, T.H. (1986) Protein Sorting in Yeast: Mutants Defective in Vacuole Biogenesis Mislocalize Vacuolar Proteins into the Late Secretory Pathway. Cell, 47, 1041-1051. http://dx.doi.org/10.1016/0092-8674(86)90819-6

[21]   Brachmann, C.B., Davies, A., Cost, G.J., Caputo, E., Li, J., Hieter, P. and Boeke, J.D. (1998) Designer Deletion Strains Derived from Saccharomyces cerevisiae S288C: A Useful Set of Strains and Plasmids for PCR-Mediated Gene Disruption and Other Applications. Yeast, 14, 115-132.

[22]   Warner, J.R. (1991) Labeling of RNA and Phosphoproteins in Saccharomyces cerevisiae. Methods in Enzymology, 194, 423-428. http://dx.doi.org/10.1016/0076-6879(91)94033-9

[23]   Lindegren, C.C. (1949) The Yeast Cell, Its Genetics and Cytology. Educational Publishers, Incorporated, St. Louis.

[24]   Robinson, J.S., Klionsky, D.J., Banta, L.M. and Emr, S.D. (1988) Protein Sorting in Saccharomyces cerevisiae: Isolation of Mutants Defective in the Delivery and Processing of Multiple Vacuolar Hydrolases. Molecular and Cellular Biology, 8, 4936-4948.

[25]   Jacobson, L., Halmann, M. and Yariv, J. (1982) The Molecular Composition of the Volutin Granule of Yeast. Biochemical Journal, 201, 473-479.

[26]   Gerrard, S.R., Bryant, N.J. and Stevens, T.H. (2000) VPS21 Controls Entry of Endocytosed and Biosynthetic Proteins into the Yeast Prevacuolar Compartment. Molecular Biology of the Cell, 11, 613-626. http://dx.doi.org/10.1091/mbc.11.2.613

[27]   Stack, J.H., Herman, P.P., Schu, P.V. and Emr, S.D. (1993) A Membrane-Associated Complex Containing the Vps15 Protein Kinase and the Vps34 PI Kinase Is Essential for Protein Sorting to the Yeast Lysosome-Like Vacuole. The EMBO Journal, 12, 2195-2204.

[28]   Piper, R.C., Cooper, A.A., Yang, H. and Stevens, T.H. (1995) VPS27 Controls Vacuolar and Endocytic Traffic through a Prevacuolar Compartment in Saccharomyces cerevisiae. The Journal of Cell Biology, 131, 603-617. http://dx.doi.org/10.1083/jcb.131.3.603

[29]   Suzuki, K. and Ohsumi, Y. (2007) Molecular Machinery of Autophagosome Formation in Yeast, Saccharomyces cerevisiae. FEBS Letters, 581, 2156-2161.
http://dx.doi.org/10.1016/j.febslet.2007.01.096

[30]   Xie, Z. and Klionsky, D.J. (2007) Autophagosome Formation: Core Machinery and Adaptations. Nature Cell Biology, 9, 1102-1109. http://dx.doi.org/10.1038/ncb1007-1102

[31]   Lynch-Day, M.A. and Klionsky, D.J. (2010) The Cvt Pathway as a Model for Selective Autophagy. FEBS Letters, 584, 1359-1366. http://dx.doi.org/10.1016/j.febslet.2010.02.013

[32]   Williams, R.L. and Urbé, S. (2007) The Emerging Shape of the ESCRT Machinery. Nature Reviews Molecular Cell Biology, 8, 355-368. http://dx.doi.org/10.1038/nrm2162

[33]   Hama, H., Tall, G.G. and Horazdovsky, B.F. (1999) Vps9p Is a Guanine Nucleotide Exchange Factor Involved in Vesicle-Mediated Vacuolar Protein Transport. The Journal of Biological Chemistry, 274, 15284-15291.

[34]   Peterson, M.R., Burd, C.G. and Emr, S.D. (1999) Vac1p Coordinates Rab and Phosphatidylinositol 3-Kinase Signaling in Vps45p-Dependent Vesicle Docking/Fusion at the Endosome. Current Biology, 9, 159-162. http://dx.doi.org/10.1016/S0960-9822(99)80071-2

[35]   Wurmser, A.E. and Emr, S.D. (2002) Novel PtdIns(3)P-Binding Protein Etf1 Functions as an Effector of the Vps34 PtdIns 3-Kinase in Autophagy. The Journal of Cell Biology, 158, 761-772.

[36]   Matsumoto, R., Suzuki, K. and Ohya, Y. (2013) Organelle Acidification Is Important for Localisation of Vacuolar Proteins in Saccharomyces cerevisiae. Protoplasma, 250, 1283-1293.
http://dx.doi.org/10.1007/s00709-013-0510-2

[37]   Tomaschevsky, A.A., Ryasanova, L.P., Kulakovskaya, T.V. and Kulaev, I.S. (2010) Inorganic Polyphosphate in the Yeast Saccharomyces cerevisiae with a Mutation Disturbing the Function of Vacuolar ATPase. Biochemistry (Moscow), 75, 1052-1054.

[38]   Rosenfeld, L., Reddi, A.R., Leung, E., Aranda, K., Jensen, L.T. and Culotta, V.C. (2010) The Effect of Phosphate Accumulation on Metal Ion Homeostasis in Saccharomyces cerevisiae. JBIC Journal of Biological Inorganic Chemistry, 15, 1051-1062. http://dx.doi.org/10.1007/s00775-010-0664-8

[39]   Ryazanova, L., Andreeva, N., Kulakovskaya, T., Valiakhmetov, A., Yashin, V., Vagabov, V. and Kulaev, I.S. (2011) The Early Stage of Polyphosphate Accumulation in Saccharomyces cerevisiae: Comparative Study by Extraction and DAPI Staining. Advances in Bioscience and Biotechnology, 2, 293-297. http://dx.doi.org/10.4236/abb.2011.24042

[40]   Puchkov, E.O. (2010) Brownian Motion of Polyphosphate Complexes in Yeast Vacuoles: Characterization by Fluorescence Microscopy with Image Analysis. Yeast, 27, 309-315.
http://dx.doi.org/10.1002/yea.1754

 
 
Top