FIG. 6.

LANA preferentially binds to the tetramerization domain of p53 in vitro. (A) Full-length p53 and the p53 transactivation domain (TR), DNA-binding domain (DBD), and tetramerization domain (TET) fused to GST were expressed in E. coli DH5α cells and purified from sonicated cell lysate with glutathione-Sepharose 4B beads (Amersham-Pharmacia). (B) Luciferase and LANA protein were in vitro transcribed and translated in the presence of [³⁵S]methionine and cysteine and incubated with equal amounts of purified p53 constructs for 4 h at 4°C. Pulled-down, in vitro-translated product was resolved on an 8% Tris-glycine gel.