PML and c-Jun colocalize and interact. (A) PML colocalizes with phosphorylated c-Jun into the UV-NB. Primary human fibroblasts were UV-treated (60 J/m2) and stained with antibodies against PML (green) and phosphorylated c-Jun (Ser 63) (P–c-Jun; red). Arrows indicate sites of PML and P–c-Jun colocalization (yellow). (B) PML interacts with c-Jun. WI-38 cells were left untreated or UV-treated (60 J/m2) and lysed after 3 hours. Anti-PML immunoprecipitates were probed with antibodies against c-Jun or PML. An isotypic IgG antibody (ISO) was used as negative control of coimmunoprecipitation. (C) c-Jun and PML directly interact. GST–c-Jun glutathione beads were incubated with in vitro–translated PML, PML RBCC (RING finger-B box-coiled coil), or PML ΔRBCC and then extensively washed. The input is 20% of the in vitro–translated PML used in the pull-down experiment. Data shown are representative of 3 independent experiments. (D) A model for PML function upon distinct genotoxic stresses. While PML regulates p53 upon γ-irradiation, it selectively modulates c-Jun proapoptotic function upon UV irradiation.
