Figure 6.

Importance of DNA-binding autoinhibition and DNA bend orientation for Stromelysin-1 promoter transactivation. (A) Detail of the structure of the Ets-1 inhibitory module around Tyr 424 (20). (B) Gel shift assays. p51Y424A was incubated with M1, M2, WT and WT + 4 variants, as defined in Table 1. Binary (2) and ternary (3) complexes are indicated. (C) Gel shift assays were performed with p51Y424A in the presence of the different probes as described in Figure 5A. (D) Relative mobility as a function of linker length expressed in base pairs and fitted to a sinusoidal function. (E) DNA-bending directions induced by p51Y424A, p42 and p51 are represented by arrows in a plane perpendicular to the long DNA axis, positioned at the EBS palindrome center. EBS 1 and 2 are the centers of the two core motifs in the WT EBS palindrome. (F) Transient transfection assays with reporter plasmids containing M1, M2, WT and WT + 4, defined in Table 1, and either p51Y424A, p51 or p42 eukaryotic expression vector. Luciferase activities were normalized and expressed as relative luciferase activity (RLU).