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

Published online by Cambridge University Press:  14 May 2008

L. D. B. DE MELO ,
C. SANT'ANNA ,
S. A. REIS ,
D. LOURENÇO ,
W. DE SOUZA ,
U. G. LOPES  and
N. L. CUNHA-E-SILVA
L. D. B. DE MELO
Affiliation:
Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brazil Departamento de Biotecnologia, Centro Federal de Educação Tecnológica de Química, Rua Senador Furtado, 121 CEP: 20270-021, Rio de Janeiro, RJ, Brazil
C. SANT'ANNA
Affiliation:
Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brazil
S. A. REIS
Affiliation:
Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brazil
D. LOURENÇO
Affiliation:
Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brazil
W. DE SOUZA
Affiliation:
Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brazil
U. G. LOPES
Affiliation:
Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brazil
N. L. CUNHA-E-SILVA*
Affiliation:
Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brazil
*
*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
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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.

Keywords

actinactin-binding proteinstrypanosomatidcytoskeleton
Type
Original Articles
Information
Parasitology , Volume 135 , Issue 8 , July 2008 , pp. 955 - 965
Copyright
Copyright © 2008 Cambridge University Press

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