Skip to main content
. Author manuscript; available in PMC: 2013 May 14.
Published in final edited form as: Cell. 2012 Aug 3;150(3):647–658. doi: 10.1016/j.cell.2012.05.045

Figure 5. Transcriptional Cooperative Systems Can Be Engineered from Weakly-Activating sTF “monomers” that Are Dimerized with a PDZ Interaction Domain.

Figure 5

(A) The dimerization interaction promotes cooperative behavior in transcriptional activation. Syntrophin PDZ domain (dark gray) was fused to the C-terminal of ZF affinity mutant 43-8-4x, and the resulting AD-carrying sTF “monomer” was expressed from ATc-inducible pGAL1. The heterologous ligand (light gray) was fused to the C-terminal of 42-10-4x, and the resulting AD-less factor was expressed from IPTG-inducible pGAL1. The factors assemble at a synthetic “dimeric” promoter to cooperatively activate downstream transcription (“IPTG+” = full induction with 20 mM IPTG).

(B) Disruption of the dimerization interaction abolishes cooperative behavior in transcriptional activation. A nonbinding ligand variant (GSGS-VKEAAA) was instead fused to 42-10-4x. Points represent mean values for three experiments ± SD.