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Evolutionary conservation of actin-binding proteins in Trypanosoma cruzi and unusual subcellular localization of the actin homologue

  • L. D. B. DE MELO (a1) (a2), C. SANT'ANNA (a1), S. A. REIS (a1), D. LOURENÇO (a1), W. DE SOUZA (a1), U. G. LOPES (a1) and N. L. CUNHA-E-SILVA (a1)...

Summary

The actin cytoskeleton controls pivotal cellular processes such as motility and cytokinesis, as well as cell-cell and cell-substrate interactions. Assembly and spatial organization of actin filaments are dynamic events regulated by a large repertoire of actin-binding proteins. This report presents the first detailed characterization of the Trypanosoma cruzi actin (TcActin). Protein sequence analysis and homology modelling revealed that the overall structure of T. cruzi actin is conserved and that the majority of amino-acid changes are concentrated on the monomer surface. Immunofluorescence assays using specific polyclonal antibody against TcActin revealed numerous rounded and punctated structures spread all over the parasitic body. No pattern differences could be found between epimastigotes and trypomastigotes or amastigotes. Moreover, in detergent extracts, TcActin was localized only in the soluble fraction, indicating its presence in the G-actin form or in short filaments dissociated from the microtubule cytoskeleton. The trypanosomatid genome was prospected to identify actin-binding and actin-related conserved proteins. The main proteins responsible for actin nucleation and treadmilling in higher eukaryotes are conserved in T. cruzi.

Copyright

Corresponding author

*Corresponding author: Laboratório de Ultraestrutura Celular Hertha Meyer, Instituto de Biofísica Carlos Chagas Filho, Av. Carlos Chagas Filho, 373, Centro de Ciências da Saúde, Bloco G – Cidade Universitária, Ilha do Fundão, CEP: 21941-902, Rio de Janeiro, RJ, Brazil. Tel: +55 21 2562 6593. Fax: +55 21 2260 2364. E-mail: narcisa@biof.ufrj.br

References

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Aiyar, A. (2000). The use of CLUSTAL W and CLUSTAL X for multiple sequence alignment. Methods in Molecular Biology 132, 221241.
Arnold, K., Bordoli, L., Kopp, J. and Schwede, T. (2006). The SWISS-MODEL workspace: a web-based environment for protein structure homology modelling. Bioinformatics 22, 195201. doi:10.1093/bioinformatics/bti770
Balcer, H. I., Goodman, A. L., Rodal, A. A., Smith, E., Kugler, J., Heuser, J. E. and Goode, B. L. (2003). Coordinated regulation of actin filament turnover by a high-molecular-weight Srv2/CAP complex, cofilin, profilin, and Aip1. Current Biology 13, 21592169. doi:10.1016/j.cub.2003.11.051
Baum, J., Papenfuss, A. T., Baum, B., Speed, T. P. and Cowman, A. F. (2006). Regulation of apicomplexan actin-based motility. Nature Reviews Microbiology 4, 621628. doi:10.1038/nrmicro1465
Bogitsh, B. J., Ribeiro-Rodrigues, R. and Carter, C. E. (1995). In vitro effects of mannan and cytochalasin B on the uptake of horseradish peroxidase and [14C] sucrose by Trypanosoma cruzi epimastigotes. Journal of Parasitology 81, 144148.
Broadhead, R., Dawe, H. R., Farr, H., Griffiths, S., Hart, S. R., Portman, N., Shaw, M. K., Ginger, M. L., Gaskell, S. J., McKean, P. G. and Gull, K. (2006). Flagellar motility is required for the viability of the bloodstream trypanosome. Nature, London 440, 224227.
Camargo, E. P. (1964). Growth and differentiation in Trypanosoma cruzi. I. Origin of metacyclic trypanosomes in liquid media. Revista do Instituto de Medicina Tropical de Sao Paulo 12, 93100.
Carlier, M. F. (1998). Control of actin dynamics. Current Opinion in Cell Biology 10, 4551. doi:10.1016/S0955-0674(98)80085-9
Cevallos, A. M., Lopez-Villasenor, I., Espinosa, N., Herrera, J. and Hernandez, R. (2003). Trypanosoma cruzi: allelic comparisons of the actin genes and analysis of their transcripts. Experimental Parasitology 103, 2734. doi:10.1016/S0014-4894(03)00066-3
Corrêa, J. R., Atella, G. C., Batista, M. M. and Soares, M. J. (2008). Transferrin uptake in Trypanosoma cruzi is impaired by interference on cytostome-associated cytoskeleton elements and stability of membrane cholesterol, but not by obstruction of clathrin-dependent endocytosis. Experimental Parasitology 119, 5866. doi:10.1016/j.exppara.2007.12.010
Cvrckova, F., Rivero, F. and Bavlnka, B. (2004). Evolutionarily conserved modules in actin nucleation: lessons from Dictyostelium discoideum and plants. Protoplasma 224, 1531.
De Melo, L. D., Nepomuceno-Silva, J. L., Sant'Anna, C., Eisele, N., Ferraro, R. B., Meyer-Fernandes, J. R., de Souza, W., Cunha-e-Silva, N. L. and Lopes, U. G. (2004). TcRho1 of Trypanosoma cruzi: role in metacyclogenesis and cellular localization. Biochemical and Biophysical Research Communications 323, 10091016. doi:10.1016/j.bbrc.2004.08.197
De Melo, L. D., Eisele, N., Nepomuceno-Silva, J. L. and Lopes, U. G. (2006). TcRho1, the Trypanosoma cruzi Rho homologue, regulates cell-adhesion properties: evidence for a conserved function. Biochemical and Biophysical Research Communications 345, 617622. doi:10.1016/j.bbrc.2006.04.075
De Sá-Freire, A., Nepomuceno-Silva, J. L., da Paixão, J. C., de Mendonça, S. M., de Melo, L. D. and Lopes, U. G. (2003). TcArf1: a Trypanosoma cruzi ADP-ribosylation factor. Parasitology Research 91, 166170. doi:10.1007/s00436-003-0952-0
De Souza, W., Meza, I., Martinez-Palomo, A., Sabanero, M., Souto-Padron, T. and Meirelles, M. N. (1983). Trypanosoma cruzi: distribution of fluorescently labeled tubulin and actin in epimastigotes. Journal of Parasitology 69, 138142.
De Souza, W. (2002). Basic cell biology of Trypanosoma cruzi. Current Pharma Design 8, 269285. doi:10.2174/1381612023396276
Disanza, A., Steffen, A., Hertzog, M., Frittoli, E., Rottner, K. and Scita, G. (2005). Actin polymerization machinery: the finish line of signaling networks, the starting point of cellular movement. Cellular and Molecular Life Science 62, 955970. doi:10.1007/s00018-004-4472-6
El-Sayed, N. M., Myler, P. J., Bartholomeu, D. C., Nilsson, D., Aggarwal, G., Tran, A. N., Ghedin, E., Worthey, E. A., Delcher, A. L., Blandin, G., Westenberger, S. J., Caler, E., Cerqueira, G. C., Branche, C., Haas, B., Anupama, A., Arner, E., Aslund, L., Attipoe, P., Bontempi, E., Bringaud, F., Burton, P., Cadag, E., Campbell, D. A., Carrington, M., Crabtree, J., Darban, H., da Silveira, J. F., de Jong, P., Edwards, K., Englund, P. T., Fazelina, G., Feldblyum, T., Ferella, M., Frasch, A. C., Gull, K., Horn, D., Hou, L., Huang, Y., Kindlund, E., Klingbeil, M., Kluge, S., Koo, H., Lacerda, D., Levin, M. J., Lorenzi, H., Louie, T., Machado, C. R., McCulloch, R., McKenna, A., Mizuno, Y., Mottram, J. C., Nelson, S., Ochaya, S., Osoegawa, K., Pai, G., Parsons, M., Pentony, M., Pettersson, U., Pop, M., Ramirez, J. L., Rinta, J., Robertson, L., Salzberg, S. L., Sanchez, D. O., Seyler, A., Sharma, R., Shetty, J., Simpson, A. J., Sisk, E., Tammi, M. T., Tarleton, R., Teixeira, S., Van Aken, S., Vogt, C., Ward, P. N., Wickstead, B., Wortman, J., White, O., Fraser, C. M., Stuart, K. D. and Andersson, B. (2005 a). The genome sequence of Trypanosoma cruzi, etiologic agent of Chagas disease. Science 309, 409415. doi:10.1126/science.1112631
El-Sayed, N. M., Myler, P. J., Blandin, G., Berriman, M., Crabtree, J., Aggarwal, G., Caler, E., Renauld, H., Worthey, E. A., Hertz-Fowler, C., Ghedin, E., Peacock, C., Bartholomeu, D. C., Haas, B. J., Tran, A. N., Wortman, J. R., Alsmark, U. C., Angiuoli, S., Anupama, A., Badger, J., Bringaud, F., Cadag, E., Carlton, J. M., Cerqueira, G. C., Creasy, T., Delcher, A. L., Djikeng, A., Embley, T. M., Hauser, C., Ivens, A. C., Kummerfeld, S. K., Pereira-Leal, J. B., Nilsson, D., Peterson, J., Salzberg, S. L., Shallom, J., Silva, J. C., Sundaram, J., Westenberger, S., White, O., Melville, S. E., Donelson, J. E., Andersson, B., Stuart, K. D. and Hall, N. (2005 b). Comparative genomics of trypanosomatid parasitic protozoa. Science 309, 404409. doi:10.1126/science.1112181
Evangelista, M., Zigmond, S. and Boone, C. (2003). Formins: signaling effectors for assembly and polarization of actin filaments. Journal of Cell Science 116, 26032611. doi:10.1242/jcs.00611
Garcia-Salcedo, J. A., Perez-Morga, D., Gijon, P., Dilbeck, V., Pays, E. and Nolan, D. P. (2004). A differential role for actin during the life cycle of Trypanosoma brucei. EMBO Journal 23, 780789. doi:10.1038/sj.emboj.7600094.
Gull, K. (1999). The cytoskeleton of trypanosomatid parasites. Annual Review of Microbiology 53, 629655. doi:10.1146/annurev.micro.53.1.629
Huang, T. Y., DerMardirossian, C. and Bokoch, G. M. (2006). Cofilin phosphatases and regulation of actin dynamics. Current Opinion in Cell Biology 18, 2631. doi:10.1016/j.ceb.2005.11.005
Kim, K., Galletta, B. J., Schmidt, K. O., Chang, F. S., Blumer, K. J. and Cooper, J. A. (2006). Actin-based motility during endocytosis in budding yeast. Molecular Biology of the Cell 17, 13541363. doi:10.1091/mbc.E05-10-0925
Lanzetti, L., Di Fiore, P. P. and Scita, G. (2001). Pathways linking endocytosis and actin cytoskeleton in mammalian cells. Experimental Cell Research 271, 4556. doi:10.1006/excr.2001.5369
Mortara, R. A. (1989). Studies on trypanosomatid actin. I. Immunochemical and biochemical identification. Journal of Protozoology 36, 813.
Nayak, R. C., Sahasrabuddhe, A. A., Bajpai, V. K. and Gupta, C. M. (2005). A novel homologue of coronin colocalizes with actin in filament-like structures in Leishmania. Molecular and Biochemical Parasitology 143, 152164. doi:10.1016/j.molbiopara.2005.06.001
Nepomuceno-Silva, J. L., Yokoyama, K., de Mello, L. D., Mendonca, S. M., Paixao, J. C., Baron, R., Faye, J. C., Buckner, F. S., Van Voorhis, W. C., Gelb, M. H. and Lopes, U. G. (2001). TcRho1, a farnesylated Rho family homologue from Trypanosoma cruzi: cloning, trans-splicing, and prenylation studies. Journal of Biological Chemistry 276, 2971129718. doi:10.1074/jbc.M102920200
Nolan, D. P. and Garcia-Salcedo, J. A. (2008). Loss of actin does not affect export of newly synthesized proteins to the surface of Trypanosoma brucei. Molecular and Biochemical Parasitology 157, 233235. doi:10.1016/j.molbiopara.2007.10.006
Ono, S. (2003). Regulation of actin filament dynamics by actin depolymerizing factor/cofilin and actin-interacting protein 1: new blades for twisted filaments. Biochemistry 42, 1336313370. doi:10.1021/bi034600x S0006-2960(03)04600-2
Paavilainen, V. O., Bertling, E., Falck, S. and Lappalainen, P. (2004). Regulation of cytoskeletal dynamics by actin-monomer-binding proteins. Trends in Cell Biology 14, 386394. doi:10.1016/j.tcb.2004.05.002
Paunola, E., Mattila, P. K. and Lappalainen, P. (2002). WH2 domain: a small, versatile adapter for actin monomers. FEBS Letters 513, 9297. doi:10.1016/S0014-5793(01)03242-2
Pollard, T. D. and Beltzner, C. C. (2002). Structure and function of the Arp2/3 complex. Current Opinion in Structural Biology 12, 768774. doi:10.1016/S0959-440(02)00396-2
Puius, Y. A., Mahoney, N. M. and Almo, S. C. (1998). The modular structure of actin-regulatory proteins. Current Opinion in Cell Biology 10, 2334. doi:10.1016/S0955-0674(98)80083-5
Sahasrabuddhe, A. A., Bajpai, V. K. and Gupta, C. M. (2004). A novel form of actin in Leishmania: molecular characterisation, subcellular localisation and association with subpellicular microtubules. Molecular and Biochemical Parasitology 134, 105114. doi:10.1016/j.molbiopara.2003.11.008
Schneider, A., Plessmann, U. and Weber, K. (1997). Subpellicular and flagellar microtubules of Trypanosoma brucei are extensively glutamylated. Journal of Cell Science 110, 431437.
Scott, V., Sherwin, T. and Gull, K. (1997). Gamma-tubulin in trypanosomes: molecular characterisation and localisation to multiple and diverse microtubule organising centres. Journal of Cell Science 110, 157168.
Sheterline, P. and Sparrow, J. C. (1994). Actin. Protein Profile 1, 1121.
Stradal, T., Kranewitter, W., Winder, S. J. and Gimona, M. (1998). CH domains revisited. FEBS Letters 431, 134137. doi:10.1016/S0014-5793(98)00751-0
Wear, M. A. and Cooper, J. A. (2004). Capping protein: new insights into mechanism and regulation. Trends in Biochemical Sciences 29, 418428. doi:10.1016/j.tibs.2004.06.003
Weaver, A. M., Young, M. E., Lee, W. L. and Cooper, J. A. (2003). Integration of signals to the Arp2/3 complex. Current Opinion in Cell Biology 15, 2330. doi:10.1016/S0955-0674(02)00015-7
Welch, M. D., DePace, A. H., Verma, S., Iwamatsu, A. and Mitchison, T. J. (1997). The human Arp2/3 complex is composed of evolutionarily conserved subunits and is localized to cellular regions of dynamic actin filament assembly. Journal of Cell Biology 138, 375384.
Wilson, W. and Seebeck, T. (1997). Identification of a profilin homologue in Trypanosoma brucei by complementation screening. Gene 187, 201209. doi:10.1016/S0378-1119(96)00749-4
Zigmond, S. H. (2004). Formin-induced nucleation of actin filaments. Current Opinion in Cell Biology 16, 99105. doi:10.1016/j.ceb.2003.10.019

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Evolutionary conservation of actin-binding proteins in Trypanosoma cruzi and unusual subcellular localization of the actin homologue

  • L. D. B. DE MELO (a1) (a2), C. SANT'ANNA (a1), S. A. REIS (a1), D. LOURENÇO (a1), W. DE SOUZA (a1), U. G. LOPES (a1) and N. L. CUNHA-E-SILVA (a1)...

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