Structural biology of the tumour suppressor p53 and its complexes
Lead Research Organisation:
MRC Laboratory of Molecular Biology
Department Name: UNLISTED
Abstract
The body has defence mechanisms against cancer that involve a protein called p53. This protein also has important roles in many other biological processes, ranging from fertility to ageing. When p53 receives signals that a cell is becoming cancerous, the protein sets into action a train of events that eventually lead to the cancer cell being destroyed. For cancer cells to survive, the activities of p53 have to be eliminated. About half of human cancers have their p53 being disabled by a mutation, most likely caused by environmental damage or radiation. In many cancers, there are raised levels of proteins that inactivate or destroy p53. Our studies examine the shapes of p53 and the other proteins and how they all fit together. These studies will provide fundamental information for designing new anti-cancer drugs that will protect p53.
Technical Summary
Intrinsically disordered domains are now known to be crucial, functional components of many proteins, especially those involved in cell signalling and regulation of the cell cycle. The presence of disordered regions prevents those proteins being crystallised for structural studies, and so solution of their structures is a major challenge, requiring techniques other than x-ray crystallography. The tumour suppressor p53 is an archetypical example of such a protein, as are its negative regulators MDM2 and MDMX. p53 is inactivated directly by mutation in ~50% of human cancers and its apoptopic pathways impaired in the remainder, often by raised levels of MDM2 and MDMX. We have recently solved the structure of p53 in solution using a combination of state-of-the-art NMR spectroscopy, small-angle-x-ray scattering, other biophysical and structural methods, protein engineering and computation. We now wish to do the same for MDMX and MDM2, and their complexes with p53. We wish to reconstruct the structure of p53 and its complexes in solution in order to understand the relationship between p53s structure and activity and how it is regulated by post-translational modification and interaction with other partner proteins. These studies will provide fundamental information on the control of the cell cycle and aid in the development of novel anti-cancer drugs. We have the necessary preliminary data to ensure that these studies should be successful.
People |
ORCID iD |
Alan Fersht (Principal Investigator) |
Publications
Natan E
(2012)
Structure and kinetic stability of the p63 tetramerization domain.
in Journal of molecular biology
Natan E
(2011)
Interaction of the p53 DNA-binding domain with its n-terminal extension modulates the stability of the p53 tetramer.
in Journal of molecular biology
Melero R
(2011)
Electron microscopy studies on the quaternary structure of p53 reveal different binding modes for p53 tetramers in complex with DNA.
in Proceedings of the National Academy of Sciences of the United States of America
Martin TG
(2016)
Design of a molecular support for cryo-EM structure determination.
in Proceedings of the National Academy of Sciences of the United States of America
Madan E
(2018)
The curcumin analog HO-3867 selectively kills cancer cells by converting mutant p53 protein to transcriptionally active wildtype p53.
in The Journal of biological chemistry
Lum JK
(2012)
Long-range modulation of chain motions within the intrinsically disordered transactivation domain of tumor suppressor p53.
in Journal of the American Chemical Society
Liu X
(2013)
Small molecule induced reactivation of mutant p53 in cancer cells
in Nucleic Acids Research
Leith JS
(2012)
Sequence-dependent sliding kinetics of p53.
in Proceedings of the National Academy of Sciences of the United States of America
Kouba T
(2019)
The Core and Holoenzyme Forms of RNA Polymerase from Mycobacterium smegmatis.
in Journal of bacteriology
Karni-Schmidt O
(2018)
Correction: Energy-dependent nucleolar localization of p53 in vitro requires two discrete regions within the p53 carboxyl terminus.
in Oncogene
Description | Support for frontier research (ERC) Advanced Grant |
Amount | € 1,459,538 (EUR) |
Funding ID | GA 268506 |
Organisation | European Research Council (ERC) |
Sector | Public |
Country | Belgium |
Start | 06/2011 |
End | 07/2017 |
Description | Electron Microscopy |
Organisation | Center for Cooperative Research in Biosciences (CIC bioGUNE) |
Country | Spain |
Sector | Charity/Non Profit |
PI Contribution | We prepared proteins and designed experiments. |
Collaborator Contribution | They performed electron microscopy experiments and analysed data. |
Impact | 17620598, 21178074 |
Start Year | 2007 |
Description | Mass Spectrometry |
Organisation | University of Oxford |
Department | Department of Chemistry |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | We did the experiments and analysis, with technical assistance from collaborators. |
Collaborator Contribution | Provided mass spectrometry facilities and intellectual input |
Impact | 19667193, 19815500, 21457718, 21525412 |
Start Year | 2009 |
Description | Single molecule studies |
Organisation | Harvard University |
Department | Department of Biological Chemistry & Molecular Pharmacology (BCMP) |
Country | United States |
Sector | Academic/University |
PI Contribution | We provided samples, did experiments and analysed data. |
Collaborator Contribution | They performed experiments and analysed data. |
Impact | 18424488, 21178072 |
Start Year | 2008 |
Title | COMPOUNDS FOR USE IN STABILIZING P53 MUTANTS |
Description | Compounds of formula (I): wherein X is selected from CRX and N; RN1 is selected from H and C1-4 alkyl, which may be substituted by SH or halo; RG1 is selected from H and SH; RC2 is selected from H and optionally substituted C1-7 alkyl; RC3 is selected from H and optionally substituted C1-7 alkyl; Rx is selected from H, OH and NH2; RC4 is selected from: (i) an optionally substituted C3-12 N-containing heterocyclyl; (ii) C(=O)NRN5RN6, where RN5 and RN6 are independently selected from H, optionally substituted C1-7 alkyl, optionally substituted C3-20 heterocyclyl and optionally substituted C5-20 aryl or RN5 and RN6 and the nitrogen atom to which they are attached form an optionally substituted N-containing C5-7 heterocyclyl group; (iii) C(=O)ORO1, where RO1 is selected from H, optionally substituted C1-7 alkyl, optionally substituted C3-20 heterocyclyl and optionally substituted C5-20 aryl; (iv) C(=O)NHNHSO2RS1, where RS1 is selected from H, optionally substituted C1-7 alkyl, optionally substituted C3-20 heterocyclyl and optionally substituted C5-20 aryl; (v) OC(=O)RC8, where RC8 is selected from H, optionally substituted C1-7 alkyl, optionally substituted C3-20 heterocyclyl and optionally substituted C5-20 aryl; (vi) OC(=O)NRN7RN8, where RN7 and RN8 are independently selected from H, optionally substituted C1-7 alkyl, optionally substituted C3-20 heterocyclyl and optionally substituted C5-20 aryl or RN7 and RN8 and the nitrogen atom to which they are attached form an optionally substituted N-containing C5-7 heterocyclyl group; and (vii) C(=O)CH2NH C(=O)NHNH2, CHC(CN)2, CHC(CN)C(=O)NH2, and carboxy; RC5 is selected from H, OH and NH2; or RC4 and RC5 together with the carbon atoms to which they are bound form an optionally substituted aromatic ring containing either 5 or 6 ring atoms, of formula: where Q represents O, N, or CRQ1=CRQ2, where RQ1 and RQ2 are independently selected from H, OH and NH2; RC6 is selected from H, OH and NH2; and RC7 is selected from optionally substituted C3-12 N-containing heterocyclyl, NHC(=O)RC9, CH2NRN2RN3 and NHC(=S)NHRN4, where RC9 is selected from optionally substituted C1-7 alkyl, optionally substituted C3-20 heterocyclyl and optionally substituted C5-20 aryl, RN2 and RN3 are independently selected from H, optionally substituted C1-7 alkyl, optionally substituted C3-20 heterocyclyl and optionally substituted C5-20 aryl or RN2 and RN3 and the nitrogen atom to which they are attached form an optionally substituted N-containing C5-7 heterocyclyl group, and RN4 is selected from optionally substituted C1-7 alkyl, optionally substituted C3-20 heterocyclyl and optionally substituted C5-20 aryl, and when RC4 and RC5 are not bound together, RC3 may additionally be selected from OR02, where RO2 is a C1-4 alkyl group, and C(=O)ORO3, where RO3 is a C1-4 alkyl group and RC2 may additionally be selected from halo, for use in stabilising a p53 protein carrying a Y220C mutation. |
IP Reference | WO2009136175 |
Protection | Patent application published |
Year Protection Granted | 2009 |
Licensed | No |
Impact | Scientific breakthrough in stabilizing p53 mutants. |
Description | Gave Cornforth lecture at Sussex University |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | National |
Primary Audience | Other academic audiences (collaborators, peers etc.) |
Results and Impact | Talk sparked questions and discussion N/A |
Year(s) Of Engagement Activity | 2013 |
Description | Gave a seminar at Parnas Meeting in Jerusalem |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Other academic audiences (collaborators, peers etc.) |
Results and Impact | Discussions N/A |
Year(s) Of Engagement Activity | 2013 |
Description | Invited Speaker at Protein Evolution Meeting in Stockholm |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Other academic audiences (collaborators, peers etc.) |
Results and Impact | Discussions N/A |
Year(s) Of Engagement Activity | 2014 |
Description | Invited speaker at Sperling Memorial Meeting, Rehovoth |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Other academic audiences (collaborators, peers etc.) |
Results and Impact | Discussion/Questions N/A |
Year(s) Of Engagement Activity | 2013 |
Description | Plenary Lecture at IUBMB, Taiwan |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Other academic audiences (collaborators, peers etc.) |
Results and Impact | Discussions N/A |
Year(s) Of Engagement Activity | 2014 |
Description | Plenary Lecture at Protein Aggregation Meeting in Umea |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Other academic audiences (collaborators, peers etc.) |
Results and Impact | Questions and discussions N/A |
Year(s) Of Engagement Activity | 2014 |
Description | Special Lecture at Trinity College Dublin |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Other academic audiences (collaborators, peers etc.) |
Results and Impact | Sparked discussions and questions N/A |
Year(s) Of Engagement Activity | 2014 |
Description | Spoke at Spetses Summer School |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | National |
Primary Audience | Other academic audiences (collaborators, peers etc.) |
Results and Impact | Discussions N/A |
Year(s) Of Engagement Activity | 2013 |