The Bub3-BD of BubR1 is necessary and sufficient for BubR1 localization on the tether. (A) Scheme of BubR1 full-length (FL) domains identified with a secondary structural prediction algorithm (globprot), and the BubR1 truncated versions along with their localization (+) or not (−) on the kinetochore (KT) or tether. The numbers correspond to the position of the first and last amino acid of the BubR1 truncation construct. The 330–762 [E481K] construct contains a substitution of E481 by K. The BubR1 constructs are fused with GFP on their N terminus. (B) Western blot of the different GFP::BubR1 constructs from transgenic adult flies. (C) Images of live neuroblasts expressing I-CreI and labeled with H2A.Z::RFP and the indicated GFP::BubR1 constructs (also see Video 1 and Video 2). The kinetochore and tether localization of the GFP::BubR1 constructs are indicated with a yellow arrow and a cyan arrowhead. The white arrowheads point to the I-CreI–induced acentric chromatids. The cells are delineated with white dotted lines. Bars, 10 µm.