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. Author manuscript; available in PMC: 2014 Jan 8.
Published in final edited form as: Science. 2013 Apr 11;340(6134):10.1126/science.1235822. doi: 10.1126/science.1235822

Fig. 2. Phosphorylation of Nud1 by Cdc15 in anaphase promotes Mob1 recruitment to SPBs.

Fig. 2

(A) Nud1-3V5 was immunoprecipitated from anaphase-arrested A31602 and A31661 cells, and bound Mob1 (IPα-HA), Nud1 (IPα-V5), Nud1 phosphorylated at T78 (IPα-pT78), total Mob1 (input α-HA), and total Nud1 (input α-V5) were examined. (B) Localization of Cdc15 and Mob1 in G1/S, metaphase, and anaphase MET3-CDC15-SPB MOB1-mCherry (A28499) cells grown in medium lacking methionine to induce expression of CDC15-SPB. (C) Phosphorylation of Nud1 T78 (α-pT78), S63 (α-pS63), S53 (α-pS53), and total Nud1 during a synchronized cell cycle (A24513). Percent meta-phase (% meta) and anaphase (% ana) cells are shown. (D) Phosphorylation of Nud1 as in Fig. 2C in anaphase-arrested NUD1-3V5 cdc14-3 (A29851) and NUD1-3V5 cdc14-3 cdc15-2 (A31661) cells. (E) Phosphorylation of Nud1 as in Fig. 2C in S-phase–arrested NUD1-3V5 (A24513) and NUD1-3V5 MET3-CDC15-SPB (A31422) cells (12). CDC15-SPB expression was induced as in Fig. 2B. (F) Phosphorylated Nud1, immunoprecipitated Cdc15 [Cdc15 IP; wild type (WT): A24957; kinase-dead (KD) Cdc15: A30371], and GST fusion protein substrate added to the kinase reaction (Substrate) are shown.