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. 2006 Dec 14;26(1):102–112. doi: 10.1038/sj.emboj.7601469

Figure 3.

Figure 3

Point mutants of MDM2 within the C-terminal tail retain the ability to homo-oligomerize through RING–RING interactions. C-terminal tail mutants in the context of MDM2 lacking the acidic domain (MDM2ΔAD) were transiently transfected into HEK293 cells together with GFP-RING (A) or GFP-RING:TY488-9AA (B). Cells were treated 36 h post-transfection with proteasome inhibitor MG132 for 4 h, lysed and immunoprecipitated with anti-MDM2 (Ab-1) and anti-GFP (7.1/13.1) antibodies, then analyzed by Western blotting. (C) Wild-type RING domains of MDM2 can homo-oligomerize in vivo with the MDM2 C-terminal tail point mutants, but not with the C-terminal tail deletion mutants. U2OS cells were cotransfected with constructs coding for GFP-tagged MDM2 RING (lacks nuclear localization signal (NLS); diffuse pattern of subcellular localization) and MDM2ΔAD (contains NLS; nuclear protein) with wild-type or mutant C-terminal tail. MDM2ΔAD-induced translocation of GFP-RING into the nucleus was used as an indicator of the interaction between the two MDM2 proteins.