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. 2017 Nov 27;114(50):E10667–E10676. doi: 10.1073/pnas.1710506114

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Copyright © 2017 the Author(s). Published by PNAS.

This open access article is distributed under Creative Commons Attribution-NonCommercial-NoDerivatives License 4.0 (CC BY-NC-ND).

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Fig. 6. — Thr28 phosphorylation in Cnp3 destroys the Dsn1-mediated interaction between Mis12C and Cnp3. (A) Sequence alignment of the two Aurora B phosphorylation sites in Dsn1 homologs (Hs, Homo sapiens; Mm, Mus musculus; Sc, Saccharomyces cerevisiae; Sp, Schizosaccharomyces pombe). Red triangles indicate two conserved serine residues that were phosphorylated by Aurora B. (B) GST pull-down assay of Mis12–Nnf1, Mis12C^WT, and Mis12C–Dsn1^DD with GST-Cnp3(1–60)^WT. (C) GST pull-down assay of Mis12C–Dsn1^DD with GST-Cnp3(1–60)^T28E. (D) The kinetics curves of Ark1 on the Dsn1 peptide (red) and the Cnp3 peptide (black) were generated by fluorometry-based kinase assay. Data were fit to the Michae-lis-Menten equation to calculate the K_m and K_cat values (Table S1).