Structural characterization of p53, c-Myc and the protein-protein interactions that regulate their biological activity
EMSL Project ID
2581
Abstract
NMR will be used to study the structure and function of (1) the tumor suppressor p53 and its interaction with the p53-regulated, pro-apoptotic protein, PIRH2; and (2) the N-terminal transactivation domain of the oncoprotein c-Myc and its interactions with several Myc interacting proteins. These projects require high-field instruments (850-900 MHz). Both projects are aimed at understanding at the molecular level, how these key growth-regulatory transcription factors (p53, c-Myc) are modulated by interactions with other proteins. PIRH2 is a newly discovered pro-apoptotic protein that is up-regulated by p53. It is also involved in a feed-back loop with p53, in that it can bind to p53 and induce the ubiquitin-dependent degradation of p53, thereby down-regulation p53 and also itself (S. Benchimol, et al, unpublished). We wish to determine the 3D structure of the 90 residue N-terminal p53-binding domain of PIRH2 and its complex with the 230 residue core domain of p53. We will need high field for improved dispersion and TROSY experiments of the complex and for improved sensitivity for PIRH2, which gives excellent HSQC spectra, but has low solubility (~0.1mM max). The N-terminal domain of c-Myc is involved in Myc-induced transcriptional activation, apoptosis, transformation and proliferation, yet very little is known at the molecular or structural level of how these activities occur. It is most likely that such activities are the result of interactions between the N-terminal domain and other regulatory proteins such as general transcription factors, kinases (Myc has several phosphorylation sites) and negative regulatory factors. We have shown that, like many transactivation domains the N-terminal domain of c-Myc is largely disordered (Pineda-Lucena et al., in preparation). We are studying complexes of this domain with two other proteins (Bin1 and TRRAP) which require high field strength for improved dispersion and sensitivity.
Project Details
Project type
Capability Research
Start Date
2002-10-01
End Date
2003-10-06
Status
Closed
Released Data Link
Team
Principal Investigator