Abstract
In order to understand the mechanism involved in Rhodotorula mucilaginosa RCL-11 resistance to copper a proteomic study was conducted. Atomic absorption spectroscopy showed that the copper concentration in the medium decreased from 0.5 to 0.19 mM 48 h after inoculation of the yeast. Analysis of one-dimensional gel electrophoresis of crude cell extracts revealed expression of differential bands between cells with and without copper. In order to study this difference, two-dimensional electrophoresis of R. mucilaginosa RCL-11 exposed to Cu for 16, 24, and 48 h was carried out. Identification of differentially expressed proteins was performed by MALDI-TOF/TOF. Ten of the 16 spots identified belonged to heat shock proteins. Superoxide dismutase, methionine synthase and beta-glucosidase were also found over-expressed at high copper concentrations. The results obtained in the present work show that when R. mucilaginosa RCL-11 is exposed to 0.5 mM copper, differential proteins, involved in cell resistance mechanisms, are expressed.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Burt ET, Daly R, Hoganson D, Tsirulnikov Y, Essmann M, Larsen B (2003) Isolation and partial characterization of Hsp90 from Candida albicans. Ann Clin Lab Sci 33:86–93
Cabiscol E, Belli G, Tamarit J, Echave P, Herrero E, Ros J (2002) Mitochondrial Hsp60, resistance to oxidative stress, and the labile iron pool are closely connected in Saccharomyces cerevisiae. J Biol Chem 277:44531–44538
Fechner LC, Gourlay-France C, Uher E, Tusseau-Vuillemin MH (2010) Adapting an enzymatic toxicity test to allow comparative evaluation of natural freshwater biofilms’ tolerance to metals. Ecotoxicology 19:1302–1311
Fridovich I (1999) Fundamental aspects of reactive oxygen species, or what’s the matter with oxygen? Ann N Y Acad Sci 893:13–18
Godon C, Lagniel G, Lee J, Buhler JM, Kieffer S, Perrot M, Boucherie H, Toledano MB, Labarre J (1998) The H2O2 stimulon in Saccharomyces cerevisiae. J Biol Chem 273:22480–22489
Gonzalez JC, Peariso K, Penner-Hahn JE, Matthews RG (1996) Cobalamin-independent methionine synthase from Escherichia coli: a zinc metalloenzyme. Biochemistry 35:12228–12234
Hondorp ER, Matthews RG (2009) Oxidation of cysteine 645 of cobalamin-independent methionine synthase causes a methionine limitation in Escherichia coli. J Bacteriol 191:3407–3410
Irazusta V, Cabiscol E, Reverter-Branchat G, Ros J, Tamarit J (2006) Manganese is the link between frataxin and iron-sulfur deficiency in the yeast model of Friedreich ataxia. J Biol Chem 281:12227–12232
Irazusta V, Obis E, Moreno-Cermeno A, Cabiscol E, Ros J, Tamarit J (2010) Yeast frataxin mutants display decreased superoxide dismutase activity crucial to promote protein oxidative damage. Free Radic Biol Med 48:411–420
Ito H, Inouhe M, Tohoyama H, Joho M (2007) Characteristics of copper tolerance in Yarrowia lipolytica. Biometals 20:773–780
Jamieson DJ (1998) Oxidative stress responses of the yeast Saccharomyces cerevisiae. Yeast 14:1511–1527
Jarosz DF, Lindquist S (2010) Hsp90 and environmental stress transform the adaptive value of natural genetic variation. Science 330:1820–1824
Jeng WY, Wang NC, Lin MH, Lin CT, Liaw YC, Chang WJ, Liu CI, Liang PH, Wang AH (2011) Structural and functional analysis of three beta-glucosidases from bacterium Clostridium cellulovorans, fungus Trichoderma reesei and termite Neotermes koshunensis. J Struct Biol 173:46–56
Ketudat Cairns JR, Esen A (2010) Beta-glucosidases. Cell Mol Life Sci 67:3389–3405
Koutmos M, Datta S, Pattridge KA, Smith JL, Matthews RG (2009) Insights into the reactivation of cobalamin-dependent methionine synthase. Proc Natl Acad Sci 106:18527–18532
Machado MD, Janssens S, Soares HM, Soares EV (2009) Removal of heavy metals using a brewer’s yeast strain of Saccharomyces cerevisiae: advantages of using dead biomass. J Appl Microbiol 106:1792–1804
Malhotra R, Singh B (2006) Ethanol-induced changes in glycolipids of Saccharomyces cerevisiae. Appl Biochem Biotechnol 128:205–213
Mapa K, Sikor M, Kudryavtsev V, Waegemann K, Kalinin S, Seidel CA, Neupert W, Lamb DC, Mokranjac D (2010) The conformational dynamics of the mitochondrial Hsp70 chaperone. Mol Cell 38:89–100
Mayer MP, Bukau B (2005) Hsp70 chaperones: cellular functions and molecular mechanism. Cell Mol Life Sci 62:670–684
Park HG, Han SI, Oh SY, Kang HS (2005) Cellular responses to mild heat stress. Cell Mol Life Sci 62:10–23
Parsell DA, Lindquist S (1993) The function of heat-shock proteins in stress tolerance: degradation and reactivation of damaged proteins. Annu Rev Genet 27:437–496
Plesofsky-Vig N, Brambl R (1998) Characterization of an 88-kDa heat shock protein of Neurospora crassa that interacts with Hsp30. J Biol Chem 273:11335–11341
Puig S, Thiele DJ (2002) Molecular mechanisms of copper uptake and distribution. Curr Opin Chem Biol 6:171–180
Raggam RB, Salzer HJ, Marth E, Heiling B, Paulitsch AH, Buzina W (2010) Molecular detection and characterisation of fungal heat shock protein 60. Mycoses. doi:10.1111/j.1439-0507.2010.01933.x
Rensing C, Fan B, Sharma R, Mitra B, Rosen BP (2000) CopA: an Escherichia coli Cu(I)-translocating P-type ATPase. Proc Natl Acad Sci U S A 97:652–656
Solioz M, Stoyanov JV (2003) Copper homeostasis in Enterococcus hirae. FEMS Microbiol Rev 27:183–195
Strain J, Lorenz CR, Bode J, Garland S, Smolen GA, Vickery LE, Culotta VC (1998) Suppressors of superoxide dismutase (SOD1) deficiency in Saccharomyces cerevisiae. Identification of proteins predicted to mediate iron-sulfur cluster assembly. J Biol Chem 273:31138–31144
Suliman HS, Sawyer GM, Appling DR, Robertus JD (2005) Purification and properties of cobalamin-independent methionine synthase from Candida albicans and Saccharomyces cerevisiae. Arch Biochem Biophys 441:56–63
Tamarit J, Cabiscol E, Aguilar J, Ros J (1997) Differential inactivation of alcohol dehydrogenase isoenzymes in Zymomonas mobilis by oxygen. J Bacteriol 179:1102–1104
Tamarit J, Irazusta V, Moreno-Cermeno A, Ros J (2006) Colorimetric assay for the quantitation of iron in yeast. Anal Biochem 351:149–151
Villegas LB, Amoroso MJ, de Figueroa LI (2005) Copper tolerant yeasts isolated from polluted area of Argentina. J Basic Microbiol 45:381–391
Villegas LB, Fernández PM, Amoroso MJ, de Figueroa LI (2008) Chromate removal by yeast isolated from sediments of a tanning factory and a mine site in Argentina. Biometals 21:591–600
Villegas LB, Amoroso MJ, de Figueroa LI (2009) Responses of Candida fukuyamaensis RCL-3 and Rhodotorula mucilaginosa RCL-11 to copper stress. J Basic Microbiol 49:395–403
Wang J, Chen C (2006) Biosorption of heavy metals by Saccharomyces cerevisiae: a review. Biotechnol Adv 24:427–451
Yoneda T, Benedetti C, Urano F, Clark SG, Harding HP, Ron D (2004) Compartment-specific perturbation of protein handling activates genes encoding mitochondrial chaperones. J Cell Sci 117:4055–4066
Zhang S, Hacham M, Panepinto J, Hu G, Shin S, Zhu X, Williamson PR (2006) The Hsp70 member, Ssa1, acts as a DNA-binding transcriptional co-activator of laccase in Cryptococcus neoformans. Mol Microbiol 62:1090–1101
Acknowledgments
This work was supported by grants from Agencia Nacional de Promoción Científica Tecnológica, FONCYT (PICT2010-0101, PICT2007-568) and Consejo de Investigaciones de la Universidad de Tucumán, CIUNT (D-415) and PIP096. We also are indebted to Dr. Jordi Tamarit (University of Lleida) for critical review of the manuscript.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Irazusta, V., Estévez, C., Amoroso, M.J. et al. Proteomic study of the yeast Rhodotorula mucilaginosa RCL-11 under copper stress. Biometals 25, 517–527 (2012). https://doi.org/10.1007/s10534-012-9531-0
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10534-012-9531-0