IBR   13079
INSTITUTO DE BIOLOGIA MOLECULAR Y CELULAR DE ROSARIO
Unidad Ejecutora - UE
artículos
Título:
Trypanosoma cruzi Bromodomain Factor 3 (TcBDF3) binds acetylated alpha-tubulin and concentrates in the flagellum during metacyclogenesis
Autor/es:
ALONSO VL; VILLANOVA VG; RITAGLIATI C; MOTTA CM; CRIBB P; SERRA EC
Revista:
EUKARYOTIC CELL
Editorial:
AMER SOC MICROBIOLOGY
Referencias:
Lugar: Washington; Año: 2014 vol. 13 p. 822 - 831
ISSN:
1535-9778
Resumen:
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 analysis (WB) and immunofluorescence (IF) 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 (PTMs), which modulate microtubules functions. Acetylated α tubulin has been found in the axoneme of flagella and cilia as well as in the subpellicular microtubules of Trypanosomatids. TcBDF3 and acetylated α tubulin partially co-localized in isolated cytoskeletons and flagella from T. cruzi epimastigotes and trypomastigotes. Interaction between the two proteins was confirmed by co-immunoprecipitation and far western blot assays with synthetic acetylated α tubulin peptides and recombinant TcBDF3.