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. 2022 Jul 1;221(8):e202108156. doi: 10.1083/jcb.202108156

Figure 2.

Figure 2.

Sgo1 makes multipartite interactions with CPC components. (A) Circular view of the EDC cross-links observed between the different subunits of the CPCISB10–280 (INCENP1–58 in yellow, Survivin in green, and Borealin10–280 in purple) and Sgo11–415 (dark blue). For clarity, only contacts between Sgo1 and the CPC subunits are shown. Intermolecular contacts of INCENP, Survivin, and Borealin with Sgo1 are shown as yellow, green, and purple lines, respectively. XiNet (Kolbowski et al., 2018) was used for data visualization. Autovalidation filter was used. (B) Cartoon representation of the crystal/nuclear magnetic resonance structures of the CPC (CPC core; PDB accession no. 2QFA; Jeyaprakash et al., 2007; Borealin dimerization domain; PDB accession no. 2KDD; Bourhis et al., 2009) and domain architecture of Sgo1 highlighting the regions involved in the CPC–Sgo1 contacts observed in A. Borealin residues in the circular view are annotated to match data deposited to the ProteomeXchange Consortium via the PRIDE repository (1–271 is equivalent to 10–280). (C and D) Representative immunofluorescence images (top) and quantification (bottom) for the analysis of the recruitment of endogenous Aurora B (C) and Borealin (D) to the LacO array in U-2 OS-LacO Haspin CM cells expressing different Sgo1-LacI-GFP constructs: LacI-GFP (n = 22 for Aurora B; n = 22 for Borealin), Sgo11–527-LacI-GFP (n = 22 for Aurora B; n = 22 for Borealin), Sgo11–130-LacI-GFP (n = 22 for Aurora B; n = 22 for Borealin), Sgo1130–280-LacI-GFP (n = 22 for Aurora B; n = 22 for Borealin), Sgo1274–415-LacI-GFP (n = 22 for Aurora B; n = 22 for Borealin), and Sgo1415–527-LacI-GFP (n = 22 for Aurora B; n = 22 for Borealin). Representative immunofluorescence images in C and D show Aurora B and Borealin signal for the same cell, thus, DAPI and GFP in C and D are the same. The graphs show the intensities of Aurora B and Borealin over GFP (dots) and the means (red bar). Data are representative of two biological replicates. Scale bar, 5 μm. One-way ANOVA with Dunnett’s multiple comparison test (****, P < 0.0001).