Structure of tumor suppressor p53 and its intrinsically disordered N-terminal transactivation domain.
Proteins with intrinsically disordered domains are implicated in a vast range of biological processes, especially in cell signaling and regulation. Having solved the quaternary structure of the folded domains in the tumor suppressor p53 by a multidisciplinary approach, we have now determined the average ensemble structure of the intrinsically disordered N-terminal transactivation domain (TAD) by using residual dipolar couplings (RDCs) from NMR spectroscopy and small-angle x-ray scattering (SAXS). Remarkably, not only were we able to measure RDCs of the isolated TAD, but we were also able to do so for the TAD in both the full-length tetrameric p53 protein and in its complex with a specific DNA response element. We determined the orientation of the TAD ensemble relative to the core domain, found that the TAD was stiffer in the proline-rich region (residues 64-92), which has a tendency to adopt a polyproline II (PPII) structure, and projected the TAD away from the core. We located the TAD in SAXS experiments on a complex between tetrameric p53 and four Taz2 domains that bind tightly to the TAD (residues 1-57) and acted as "reporters." The p53-Taz2 complex was an extended cross-shaped structure. The quality of the SAXS data enabled us to model the disordered termini and the folded domains in the complex with DNA. The core domains enveloped the response element in the center of the molecule, with the Taz2-bound TADs projecting outward from the core.
Wells
Mark
M
MRC Centre for Protein Engineering, Hills Road, Cambridge CB2 0QH, United Kingdom.
Tidow
Henning
H
Rutherford
Trevor J
TJ
Markwick
Phineus
P
Jensen
Malene Ringkjobing
MR
Mylonas
Efstratios
E
Svergun
Dmitri I
DI
Blackledge
Martin
M
Fersht
Alan R
AR
D020816:Amino Acid Motifs; D004247:DNA / Q000737:chemistry; D006801:Humans; D009154:Mutation; D010446:Peptide Fragments; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D020218:Response Elements; D053838:Scattering, Small Angle; D015533:Transcriptional Activation; D016159:Tumor Suppressor Protein p53 / Q000737:chemistry*; D014961:X-Ray Diffraction
D010446:Peptide Fragments; D016159:Tumor Suppressor Protein p53; D004247:DNA