Figure 5. SUMO ligase activity of TRIM11, and functional cooperation of TRIM11’s discrete activities in the clearance of Atxn1 82Q.
(A and B) SUMOylation of HA-Atxn1 82Q by Flag-TRIM proteins in the presence of 6xHis-SUMO2 in HEK293T cells.
(C) SUMOylation of Flag-Atxn1 82Q by HA-TRIM11 in HEK293T cells treated with control or SUMO2/3 siRNA.
(D) SUMOylation of Flag-Atxnl 82Q, but not α-Syn, by HA-TRIM11 in the presence of 6xHis-SUMO2 in HEK293T cells stably expressing α-Syn.
(E) Levels of GFP-Atxn1 82Q when expressed alone or together with Flag-TRIM11/TRIM112EA in HCT116 cells.
(F) Amyloid contents in HCT116 cells transfected with TRIM11 or TRIM112EA
(G) Half-life of Flag-Atxn1 82Q in HEK293T cells when co-expressed with the indicated TRIM11 proteins. Relative Atxn1 82Q/GAPDH ratios are shown. The exposure of different blots was adjusted so that band intensities at time 0 were comparable.
Data are mean ± SD (n = 4 biological replicates). **P < 0.01.