FIGURE 5.

SIRT2 binds and deacetylates TUG. A, lysates from 293 cells transfected with FLAG-tagged SIRT2 were subjected to immunoprecipitation (IP) using beads with or without a cross-linked anti-TUG antibody, as indicated. Eluted proteins were immunoblotted (WB) to detect SIRT2 and TUG, as indicated. The experiment was repeated at least twice with consistent results. B, endogenous TUG proteins were immunoprecipitated from untransfected or SIRT2-transfected 293 cells. Eluted proteins were immunoblotted to detect AcK. The experiment was repeated twice with consistent results. C, synthetic peptides corresponding to TUG(528–550), with AcK549, or p53(368–393), with AcK382, were incubated in the presence of recombinant SIRT2 or SIRT1. All peptides had an N-terminal biotin. After incubation for the indicated times, peptides were spotted on nitrocellulose membranes and immunoblotted to detect AcK or total biotinylated peptide. Experiments were repeated three times with similar results.