Abstract
Trypanosoma cruzi is the etiological agent of Chagas disease, a major health problem in Latin America. Polyamines are polycationic compounds that play a critical role as regulators of cell growth and differentiation. In contrast with other protozoa, T. cruzi is auxotrophic for polyamines because of its inability to synthesize putrescine due to the lack of both, arginine and ornithine decarboxylase; therefore, the intracellular availability of polyamines depends exclusively on transport processes. In this work, the polyamine transporter TcPAT12 was overexpressed in T. cruzi epimastigotes demonstrating that growth rates at different concentrations of polyamines strongly depend on the regulation of the polyamine transport. In addition, parasites overexpressing TcPAT12 showed a highly increased resistance to hydrogen peroxide and the trypanocidal drugs nifurtimox and benznidazole, which act by oxidative stress and interfering the synthesis of polyamine derivatives, respectively. Finally, the presence of putative polyamine transporters was analyzed in T. cruzi, Trypanosoma brucei, and Leishmania major genomes identifying 3–6 genes in these trypanosomatids.
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Aiyar A (2000) The use of CLUSTAL W and CLUSTAL X for multiple sequence alignment. Methods Mol Biol 132:221–241
Alcazar R, Tiburcio AF (2014) Plant polyamines in stress and development: an emerging area of research in plant sciences. Front Plant Sci 5:319
Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ (1990) Basic local alignment search tool. J Mol Biol 215:403–410
Bacchi CJ, Vergara C, Garofalo J, Lipschik GY, Hutner SH (1979) Synthesis and content of polyamines in bloodstream Trypanosma brucei. J Protozool 26:484–488
Bacchi CJ, Nathan HC, Hutner SH, McCann PP, Sjoerdsma A (1980) Polyamine metabolism: a potential therapeutic target in trypanosomes. Science 210:332–334
Bachrach U, Brem S, Wertman SB, Schnur LF, Greenblatt CL (1979) Leishmania spp.: cellular levels and synthesis of polyamines during growth cycles. Exp Parasitol 48:457–463
Bailey TL, Williams N, Misleh C, Li WW (2006) MEME: discovering and analyzing DNA and protein sequence motifs. Nucl Acids Res 34:W369–W373
Barrett MP, Burchmore RJ, Stich A, Lazzari JO, Frasch AC, Cazzulo JJ, Krishna S (2003) The trypanosomiases. Lancet 362:1469–1480
Bouvier LA, Silber AM, Galvao Lopes C, Canepa GE, Miranda MR, Tonelli RR, Colli W, Alves MJ, Pereira CA (2004) Post genomic analysis of permeases from the amino acid/auxin family in protozoan parasites. Biochem Biophys Res Commun 321:547–556
Brun R, Schonenberger M (1979) Cultivation and in vitro cloning or procyclic culture forms of Trypanosoma brucei in a semi-defined medium. Short communication. Acta Trop 36:289–292
Camargo EP (1964) Growth and differentiation in Trypanosoma Cruzi. I. Origin of metacyclic trypanosomes in liquid media. Rev Inst Med Trop Sao Paulo 6:93–100
Carrillo C, Cejas S, Gonzalez NS, Algranati ID (1999) Trypanosoma cruzi epimastigotes lack ornithine decarboxylase but can express a foreign gene encoding this enzyme. FEBS Lett 454:192–196
Carrillo C, Cejas S, Huber A, Gonzalez NS, Algranati ID (2003) Lack of arginine decarboxylase in Trypanosoma cruzi epimastigotes. J Eukaryot Microbiol 50:312–316
Carrillo C, Canepa GE, Algranati ID, Pereira CA (2006) Molecular and functional characterization of a spermidine transporter (TcPAT12) from Trypanosoma cruzi. Biochem Biophys Res Commun 344:936–940
Chagas C (1909) Nova Tripanosomiaze Humana: estudos sobre amorfolojia e o ciclo evolutivo do Schizotrypanum cruzi n. gen., n. sp., ajente etiolojico de uma nova entidade mórbida do homem. Mem Inst Oswaldo Cruz 1:159–218
Colotti G, Ilari A (2011) Polyamine metabolism in Leishmania: from arginine to trypanothione. Amino Acids 40:269–285
Cupello MP, Souza CF, Buchensky C, Soares JB, Laranja GA, Coelho MG, Cricco JA, Paes MC (2011) The heme uptake process in Trypanosoma cruzi epimastigotes is inhibited by heme analogues and by inhibitors of ABC transporters. Acta Trop 120:211–218
Fairlamb AH, Blackburn P, Ulrich P, Chait BT, Cerami A (1985) Trypanothione: a novel bis(glutathionyl)spermidine cofactor for glutathione reductase in trypanosomatids. Science 227:1485–1487
Hall BS, Bot C, Wilkinson SR (2011) Nifurtimox activation by trypanosomal type I nitroreductases generates cytotoxic nitrile metabolites. J Biol Chem 286:13088–13095
Hasne MP, Coppens I, Soysa R, Ullman B (2010) A high-affinity putrescine-cadaverine transporter from Trypanosoma cruzi. Mol Microbiol 76:78–91
Igarashi K, Kashiwagi K (2000) Polyamines: mysterious modulators of cellular functions. Biochem Biophys Res Commun 271:559–564
Magnes C, Fauland A, Gander E, Narath S, Ratzer M, Eisenberg T, Madeo F, Pieber T, Sinner F (2014) Polyamines in biological samples: rapid and robust quantification by solid-phase extraction online-coupled to liquid chromatography-tandem mass spectrometry. J Chromatogr A 1331:44–51
Page RD (1996) TreeView: an application to display phylogenetic trees on personal computers. Comput Appl Biosci 12:357–358
Pereira CA, Alonso GD, Paveto MC, Flawia MM, Torres HN (1999) L-arginine uptake and L-phosphoarginine synthesis in Trypanosoma cruzi. J Eukaryot Microbiol 46:566–570
Pereira CA, Alonso GD, Ivaldi S, Silber AM, Alves MJ, Torres HN, Flawia MM (2003) Arginine kinase overexpression improves Trypanosoma cruzi survival capability. FEBS Lett 554:201–205
Rassi A Jr, Rassi A, Marin-Neto JA (2010) Chagas disease. Lancet 375:1388–1402
Tamura K, Stecher G, Peterson D, Filipski A, Kumar S (2013) MEGA6: molecular evolutionary genetics analysis version 6.0. Mol Biol Evol 30:2725–2729
Trochine A, Creek DJ, Faral-Tello P, Barrett MP, Robello C (2014) Benznidazole biotransformation and multiple targets in Trypanosoma cruzi revealed by metabolomics. PLoS Negl Trop Dis 8:e2844
Vazquez MP, Levin MJ (1999) Functional analysis of the intergenic regions of TcP2beta gene loci allowed the construction of an improved Trypanosoma cruzi expression vector. Gene 239:217–225
Young GB, Jack DL, Smith DW, Saier MH Jr (1999) The amino acid/auxin:proton symport permease family. Biochim Biophys Acta 1415:306–322
Acknowledgments
Special thanks to Lic. Fabio di Girolamo (IDIM-CONICET) for technical support. This work was supported by Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET, PIP 2011-0263, and 2013-0664), Agencia Nacional de Promoción Científica y Tecnológica (FONCYT PICT 2012-0559 and 2013-2218). CAP and MRM are members of the career of scientific investigator; CR, MS, and EVV are research fellows from CONICET.
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Reigada, C., Sayé, M., Vera, E.V. et al. Trypanosoma cruzi Polyamine Transporter: Its Role on Parasite Growth and Survival Under Stress Conditions. J Membrane Biol 249, 475–481 (2016). https://doi.org/10.1007/s00232-016-9888-z
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DOI: https://doi.org/10.1007/s00232-016-9888-z