FIGURE 4:
Targeting of Tip60 to centromeres dysregulates the localization of the Aurora B and other proteins of the Centromere Signaling Network (CSN). (A) Aurora B at centromeres is reduced when Tip60 is targeted to centromeres. Cells were arrested in mitosis with nocodazole and stained for the indicated antibody. CB-GFP n = (44, 16); CB-Tip60 n = (43, 13); p value = (0.0015; 0.033); scale bar, 5 µm. (B) Tip60 targeting affects many kinetochore/inner centromere events in an acetyltransferase dependent manner. Cells expressing CB-Tip60 were treated with either DMSO or the Tip60 acetyltransferase inhibitor Nu9056, and the localization of the indicated protein or PTM were imaged by immunofluorescence. Light and dark blue dots indicate two independent experimental replicates. Quantification of fluorescence signal of Aurora B nDMSO = (10, 9), nNU9056 = (11, 10), p value = (0.01, 0.0034); Sgo1 nDMSO = (10, 10), nNU9056 = (9, 11), p value = (0.049, 0.042); CENP-T nDMSO = (6, 11), nNU9056 = (10, 11), p value = (0.486, 0.051); Bub1 nDMSO = (11, 10), nNU9056 = (12, 11); p value = (0.4184, 0.6519); histone H3T3ph nDMSO = (8, 9), nNU9056 = (7, 9); p value (0.0018, 0.0025) and histone H2AT120ph nDMSO = (5, 9), nNU9056 = (6, 11) p value = (0.029, 0.0017) at centromeres in cells with Tip60 targeted to centromeres by CENP-B fusion and treated with Nu9016. For statistical analysis Welch’s t test was applied. ***P < 0.001, **P < 0.01, *P < 0.05, and ns P > 0.05.