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. 2011 Feb 4;30(6):1054–1066. doi: 10.1038/emboj.2011.23

Figure 1.

Figure 1

MYBBP1A is necessary for p53 acetylation induced by nucleolar disruption. (A) Knockdown of TIF-IA induces acetylation at multiple lysine residues in p53. MCF-7 cells were treated with siCont or siTIF-IA for 48 h, and the cell lysates were analysed by immunoblot using the indicated antibodies. (B) Nucleolar disruption releases the nucleolar protein MYBBP1A into the nucleoplasm. MCF-7 cells were treated with siCont or siTIF-IA for 48 h, and the localization of MYBBP1A and UBF was visualized by immunofluorescence using anti-MYBBP1A and anti-UBF antibodies. (C) Knockdown of MYBBP1A antagonizes the acetylation of p53 proteins caused by nucleolar disruption. MCF-7 cells were treated with the siCont or MYBBP1A siRNA (siMYBBP) with or without siTIF-IA for 48 h, and cell lysates were analysed by immunoblot using the indicated antibodies. (D) Knockdown of MYBBP1A decreases the acetylation levels at multiple lysine residues in the p53 protein. (Left) MCF-7 cells were treated with the siCont or two independent siMYBBPs (siMYBBP and siMYBBP-2) with or without siTIF-IA for 48 h, and cell lysates were analysed by immunoblot using site-specific acetylated p53 antibodies. (Right) Relative quantification of acetylation levels of the p53 protein. The intensities of the acetylated p53 proteins were corrected using the p53 protein level. The intensity of the siCont-treated cells was normalized to 1.0. (E) Knockdown of MYBBP1A antagonizes the acetylation of p53 induced by low-dose ActD treatment. MCF-7 cells were treated with siCont or siMYBBP for 48 h, followed by 5 nM ActD treatment for indicated times. The cell lysates were analysed by immunoblot using the indicated antibodies. (F) Recovery of the acetylation levels of p53 protein by introducing siRNA-resistant MYBBP1A along with siRNA for MYBBP1A. p53-deficient H1299 cells were treated with the siCont or siMYBBP with or without siTIF-IA for 24 h before transfection with Myc-p53 and siRNA-resistant MYBBP1A (MYBBP1A-simut). Twenty-four hours after transfection, cell lysates were analysed by immunoblot with the indicated antibodies.