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. 2010 Jan 25;285(15):11458–11464. doi: 10.1074/jbc.M109.087585

FIGURE 4.

FIGURE 4.

SIRT1 binds to acetylated TIP60. A, TIP60 specifically interacts with SIRT1 in vivo. 293T cells were transfected with plasmids encoding each of the seven Myc-tagged SIRTs (SIRT1 to -7), together with plasmids encoding SFB-TIP60. Cell extracts were precipitated (IP) using S protein-Sepharose beads. The precipitates were analyzed by immunoblotting (WB) using antibodies as indicated. B, a co-immunoprecipitation assay was performed to verify the interaction between TIP60 and SIRT1. C, SIRT1 failed to bind to unacetylated TIP60. 293T cells were transfected with plasmids encoding wild type or the HAT mutant of SFB-TIP60. Cell extracts were precipitated with S protein-Sepharose beads and immunoblotted using antibodies as indicated. D, recombinant MBP-TIP60 was autoacetylated to different levels in vitro before they were used in a pull-down assay and incubated with U2OS cell extracts. Inputs and MBP-TIP60-associated SIRT1 were analyzed by immunoblotting using indicated antibodies.