Antiproliferative Factor-Induced Changes in Phosphorylation and Palmitoylation of Cytoskeleton-Associated Protein-4 Regulate Its Nuclear Translocation and DNA Binding
Figure 7
CKAP4 binds directly to genomic DNA. (a) CKAP4 binds gDNA in an APF-dependent manner (lane 3) when compared to mock-treated control (lane 4). The CKAP4-gDNA binding is phosphorylation dependent as CKAP4 isolated from cells treated without phosphatase inhibitors prior to APF treatment failed to bind gDNA (lane 6). As controls, we loaded APF treated nuclear lysates in lane 1 and phosphatase/APF treated lysates in lane 5. Lane 2 is a no gDNA cellulose control. (b) The ability of transiently transfected V5-tagged, CKAP4 C100S/SΔE to bind gDNA in the absence of APF treatment (c) and the ability of purified rCKAP4 (residues 126–501, which includes the bZIP-like DNA-binding domain) to bind gDNA were also assessed by comparing the amount of CKAP4 captured relative to what remained in the cell lysate after binding (Sup).
