Hot spot mutp53 proteins and R248W preferential binding to scDNA.

<p>A) Distribution of hot spot mutations on full length p53 (conserved regions marked by cylinders) with a bar code representing their frequency; scheme of p53 molecules (p53 (aa 1–393), p53CΔ30 (aa 1–363)), DNA binding domains (shaded, central (CD) and C-terminal (CTDBS)) and positions of mAb epitope of DO1 (aa 21–25) are marked. The conformation of mutp53 proteins is labeled: grey (conformation mutants) and black (contact mutants). B) ScDNA binding of p53 R248W by agarose electrophoresis and DO1 immunoblotting. Increasing amounts of p53 protein (marked by the p53/DNA molar ratio) (lanes 2–5) were incubated with scDNA (pBSK, 200 ng) for 20 min and then separated in a 1% agarose gel at 4°C. DO1 immunodetection of R248W binding to scDNA showed that each retarded band of Et-Br visualized DNA on the agarose gel (lanes 2–5) corresponds to a p53 band on the DO1 immunoblot (lanes 7–10). Open circle DNA (oc) was not bound by p53.