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Fig. 1. T. brucei H2AZ localizes to distinct foci within the nucleus. (A) Sequence conservation of H2AZ. For each H2AZ, the polypeptide length is denoted as well as the percent sequence identity/similarity relative to T. brucei H2AZ. The amino terminal extension of T. brucei H2AZ (red) is unique and was excluded from these calculations. (B) A polyclonal antibody to a 14 amino-acid region of the amino-terminal tail of H2AZ (marked by a black bar in A) recognizes endogenous H2AZ in wild-type procyclic (PF) and bloodstream-form (BF) cell lines as well as those in which ectopically expressed GFP- or TY1-tagged H2AZ has replaced the endogenous H2AZ. (C) A comparison of the localization of H2AZ (red) and TY1-H2A (green) by indirect immunofluorescence in a bloodstream-form cell line over the course of the cell cycle reveals that H2AZ is not uniformly distributed. Both nuclear (n) and mitochondrial kinetoplast (k) DNA (blue) were detected with DAPI. Owing to its compact state, the kinetoplast DNA signal appears stronger than the nuclear DNA at this focal plane. Top, an interphase cell (1n 1k); middle, an early mitotic cell (1n 2k); bottom, cytokinesis (2n 2k). Bar, 2 µm.





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