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. 2014 Sep 23;289(45):31617–31623. doi: 10.1074/jbc.M114.583542

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© 2014 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 1. — Schematic of our experimental system. A, conventional apoptotic pathway. B, hypothesis: Does rapid depletion of ICAD-L trigger CAD activation and cell death? C, experimental system using AGI:TIR1 cells. A plasmid encoding OsTIR1 and AID-GFP-mICAD-L was transfected into ICAD^−/−CAD^+/+ chicken DT40 cells. F-box protein OsTIR1 binds to endogenous Skp1 to form the SCF^TIR1(Skp1-Cullin1-TIR1) complex. TIR1 binds to an AID tag in the presence of auxin (27). SCF^TIR1 ubiquitinates the AID tag and promotes the degradation of AID-tagged ICAD protein by proteasome. We set out to test whether rapid degradation of ICAD could activate CAD.