Figure 5
From: SUMOylation of ZFP282 potentiates its positive effect on estrogen signaling in breast tumorigenesis
ZFP282 is covalently modified by SUMO. (a, b) In vivo SUMOylation assay. COS-7 cells were transfected with pSG5.HA-ZFP282 and a plasmid encoding EGFP-SUMO1 as indicated. Cell extracts were prepared in the presence of N-ethylmaleimide and analyzed by immunoblot with anti-HA antibody (a) or by immunoprecipitation and immunoblot with the indicated antibodies (b). (c) SUMOylation of ZFP282 in E. coli. GST-ZFP282 was expressed in E. coli in the presence or absence of pT-E1E2S1 plasmid encoding Aos1/Uba2, Ubc9, and SUMO1. The purified ZFP282 proteins were analyzed by SDS–PAGE followed by Coomassie blue staining. (d) COS-7 cells were transfected with pSG5.HA-ZFP282 deletion mutants and a plasmid encoding EGFP-SUMO1 as indicated, and immunoblot was performed as in (a). (e) COS-7 cells were transfected with a plasmid encoding EGFP-SUMO1 and pSG5.HA-ZFP282 WT or various KR mutants as indicated, and immunoblot was performed as in (a).
