Figure 1
From: TATA binding protein associated factor 3 (TAF3) interacts with p53 and inhibits its function
Dmp53 interacts with DmTAF3. (A) Full length DmTAF3 and the clones identified from Y2H screen as p53 interacting proteins (clone 5 and clone 11) are depicted. HFD, histone fold domain; PHD, plant homeodomain. (B) The various portions of Dmp53 and human p53 that were fused to lexA-DBD and used in Y2H experiments are depicted. TA, transactivation domain; DBD, DNA binding domain; O, oligomerization domain; B, basic regulatory domain. (C) The indicated lexA-DBD fusion constructs were co-introduced into yeast with DmTAF3 amino acids 514 to 924 fused to Gal4 activation domain (AD). Interacting proteins result in complementation of His-auxotrophy and induction of β-galactosidase activity, which was detected by filter assay. Negative control was DBD-ADA2A with the same DmTAF3 clone. Positive control was DBD-rpb4 with AD-rpb7. The p53ΔN-DmTAF3 interaction was examined on a separate filter with the same positive and negative controls, and the image was fitted into the figure. (D) Direct binding of DmTAF3 to Dmp53C in vitro. Coomassie stained gel of bacterially expressed GST (lane 1) and GST-Dmp53C (lane 2). DmTAF3 (aa 514–924) was expressed in vitro in the presence of 3H-labelled leucine and incubated with GST (negative control) or GST-Dmp53C. Glutathione-agarose bead bound proteins were eluted, electrophoretically separated and detected by autoradiography (lane3 and 4).