title: Trypanosoma cruzi bromodomain factor 3 binds acetylated α-tubulin and concentrates in the flagellum during metacyclogenesis.
abstract: Bromodomains are highly conserved acetyl-lysine binding domains found mainly in proteins associated with chromatin and nuclear acetyltransferases. The Trypanosoma cruzi genome encodes at least four bromodomain factors (TcBDFs). We describe here bromodomain factor 3 (TcBDF3), a bromodomain-containing protein localized in the cytoplasm. TcBDF3 cytolocalization was determined, using purified antibodies, by Western blot and immunofluorescence analyses in all life cycle stages of T. cruzi. In epimastigotes and amastigotes, it was detected in the cytoplasm, the flagellum, and the flagellar pocket, and in trypomastigotes only in the flagellum. Subcellular localization of TcBDF3 was also determined by digitonin extraction, ultrastructural immunocytochemistry, and expression of TcBDF3 fused to cyan fluorescent protein (CFP). Tubulin can acquire different posttranslational modifications, which modulate microtubule functions. Acetylated α-tubulin has been found in the axonemes of flagella and cilia, as well as in the subpellicular microtubules of trypanosomatids. TcBDF3 and acetylated α-tubulin partially colocalized in isolated cytoskeletons and flagella from T. cruzi epimastigotes and trypomastigotes. Interaction between the two proteins was confirmed by coimmunoprecipitation and far-Western blot assays with synthetic acetylated α-tubulin peptides and recombinant TcBDF3.
authors.affiliation: Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario (UNR). Rosario, Argentina Instituto de Biología Molecular y Celular de Rosario (IBR), CONICET, Rosario, Argentina.
authors.forename: Victoria Lucia
authors.initials: VL
authors.lastname: Alonso
authors.affiliation: Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario (UNR). Rosario, Argentina Instituto de Biología Molecular y Celular de Rosario (IBR), CONICET, Rosario, Argentina.
authors.forename: Gabriela Vanina
authors.initials: GV
authors.lastname: Villanova
authors.affiliation: Instituto de Biología Molecular y Celular de Rosario (IBR), CONICET, Rosario, Argentina.
authors.forename: Carla
authors.initials: C
authors.lastname: Ritagliati
authors.affiliation: Laboratório de Ultraestrutura Celular Hertha Meyer, Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil.
authors.forename: María Cristina
authors.initials: MC
authors.lastname: Machado Motta
authors.affiliation: Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario (UNR). Rosario, Argentina Instituto de Biología Molecular y Celular de Rosario (IBR), CONICET, Rosario, Argentina.
authors.forename: Pamela
authors.initials: P
authors.lastname: Cribb
authors.affiliation: Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario (UNR). Rosario, Argentina Instituto de Biología Molecular y Celular de Rosario (IBR), CONICET, Rosario, Argentina eserra@fbioyf.unr.edu.ar.
authors.forename: Esteban Carlos
authors.initials: EC
authors.lastname: Serra
mesh_terms: D000107:Acetylation; D003593:Cytoplasm / Q000378:metabolism; D005407:Flagella / Q000378:metabolism* / Q000648:ultrastructure; D008018:Life Cycle Stages; D008870:Microtubules / Q000378:metabolism; D011485:Protein Binding; D011499:Protein Processing, Post-Translational; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D015800:Protozoan Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D014157:Transcription Factors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D014349:Trypanosoma cruzi / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D014404:Tubulin / Q000378:metabolism*
chemical_list: D015800:Protozoan Proteins; D014157:Transcription Factors; D014404:Tubulin
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