Evolutionary conservation of actin-binding proteins in Trypanosoma cruzi and unusual subcellular localization of the actin homologue.
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.
De Melo
L D B
LD
Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brazil.
Sant'Anna
C
C
Reis
S A
SA
Lourenço
D
D
De Souza
W
W
Lopes
U G
UG
Cunha-e-Silva
N L
NL
D000199:Actins / Q000032:analysis / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D000818:Animals; D000913:Antibodies, Protozoan; D000918:Antibody Specificity; D017124:Conserved Sequence; D019143:Evolution, Molecular; D006801:Humans; D042541:Intracellular Space; D008840:Microfilament Proteins / Q000737:chemistry* / Q000235:genetics*; D008969:Molecular Sequence Data; D017434:Protein Structure, Tertiary; D014349:Trypanosoma cruzi / Q000235:genetics / Q000276:immunology / Q000378:metabolism*
D000199:Actins; D000913:Antibodies, Protozoan; D008840:Microfilament Proteins