XB-IMG-168948
Xenbase Image ID: 168948
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Figure 4. Cdk1 phosphorylation of the linkers of C2H2 zinc finger proteins.(a) Linker sequences of (human) Ikaros, Sp1 and YY1. For each linker, the phosphoacceptor Ser or Thr is shown in parentheses and printed in blue in the sequence, while the Arg/Lys in the +3 position in red. (b) GST-fused linker-spanning peptides (WT or a Ser/Thr → Ala mutant; for the residues of the peptides, see Methods) of the indicated zinc finger proteins were subjected to in vitro kinase assays by using [Υ-32P]ATP and either Cdk1, Plk1 or Aurora B. For each linker peptide of each protein, relative intensity of the radioactive signal (WT = 100) is shown at the bottom. (c)–(e) In vitro Cdk1 kinase assays of the linker-spanning peptides (WT or the indicated Ala-mutants) of Ikaros (c), Sp1 (d) and YY1 (e). (f) Flag/His6-tagged YY1 protein (WT or T348A) was incubated with either interphase (I) or M phase (M) extracts (from Xenopus eggs) in the presence or absence of roscovitine (Ros; 300 μM), BI2536 (BI; 15 μM) or ZM447439 (ZM; 30 μM), and then subjected to Flag-immunoprecipitation followed by immunoblotting with the indicated antibodies. In input, the arrowhead shows Flag/His6-tagged YY1(WT). Full scans of all autoradiographies and immunoblots are included in Supplementary Fig. S5. Image published in: Suzuki K et al. (2015) Copyright © 2015, Macmillan Publishers Limited. Creative Commons Attribution-NonCommercial-ShareAlike license Larger Image Printer Friendly View |