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. 2004 Jul;24(13):6021–6028. doi: 10.1128/MCB.24.13.6021-6028.2004

FIG. 5.

FIG. 5.

SENP1 desumoylates HDAC1 and inhibits HDAC1 repressive activity. (A) SENP1 physically interacts with HDAC1 in vivo. Precipitates with glutathione-Sepharose (top) or whole-cell lysates (bottom) from COS-7 cells transfected with the indicated expression plasmid were analyzed by Western blotting with anti-Flag M2. (B) SENP1 deconjugates SUMO-1 from HDAC1 in vivo. Whole-cell lysates from COS-7 cells transfected with the indicated expression plasmids were immunoprecipitated with the anti-Flag M2 antibody and analyzed by Western blotting with the anti-HA antibody (top) or anti-HDAC1 antibody (bottom). (C and D) SENP1 overcomes HDAC1's repressive activity. PC-3 cells were transfected with Gal4-luciferase (100 ng) and either Gal4-DBD, Gal4-DBD-HDAC1, or Gal4-DBD-HDAC1 DM (100 ng) in the absence or presence of wild-type SENP1 or mutant SENP1 (SENP1mut) plasmids (150 ng). The luciferase activity was measured. (E) SENP1 reduces the deacetylase activity of HDAC1. HDAC1 or HDAC1 DM was immunoprecipitated by the anti-Flag M2 antibody from the nuclear extracts of HeLa cells transfected with Flag-HDAC1, Flag-HDAC1 DM, or Flag-HDAC1 plus either SENP1 or mutant SENP1 plasmids and assayed for HDAC activity. The immunoprecipitates were quantified by Western blotting with the anti-HDAC1 antibody.