Structural properties of the promiscuous VP16 activation domain.
Herpes simplex virion protein 16 (VP16) contains two strong activation regions that can independently and cooperatively activate transcription in vivo. We have identified the regions and residues involved in the interaction with the human transcriptional coactivator positive cofactor 4 (PC4) and the general transcription factor TFIIB. NMR and biochemical experiments revealed that both VP16 activation regions are required for the interaction and undergo a conformational transition from random coil to alpha-helix upon binding to its target PC4. The interaction is strongly electrostatically driven and the binding to PC4 is enhanced by the presence of its amino-terminal domain. We propose models for binding of VP16 to the core domains of PC4 and TFIIB that are based on two independent docking approaches using NMR chemical shift changes observed in titration experiments. The models are consistent with results from site-directed mutagenesis and provide an explanation for the contribution of both acidic and hydrophobic residues for transcriptional activation by VP16. Both intrinsically unstructured activation domains are attracted to their interaction partner by electrostatic interactions, and adopt an alpha-helical conformation around the important hydrophobic residues. The models showed multiple distinct binding surfaces upon interaction with various partners, providing an explanation for the promiscuous properties, cooperativity, and the high activity of this activation domain.
Jonker
Hendrik R A
HR
Bijvoet Center for Biomolecular Research, section NMR spectroscopy, Utrecht University, Padualaan 8, 3584 CH, Utrecht, The Netherlands.
Wechselberger
Rainer W
RW
Boelens
Rolf
R
Folkers
Gert E
GE
Kaptein
Rob
R
D017405:Herpes Simplex Virus Protein Vmw65 / Q000737:chemistry / Q000378:metabolism*; D008958:Models, Molecular; D019906:Nuclear Magnetic Resonance, Biomolecular; D011485:Protein Binding; D011487:Protein Conformation; D055672:Static Electricity; D035581:Transcription Factor TFIIB / Q000378:metabolism
D017405:Herpes Simplex Virus Protein Vmw65; D035581:Transcription Factor TFIIB