1. The crystal structure of the R280K mutant of human p53 explains the loss of DNA binding
- Author
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Benedita A. Pinheiro, Sara Gomes, Maria João Romão, Lucília Domingues, Ana Sara Gomes, Filipe Freire, Lucília Saraiva, Ana Luísa Carvalho, Carla Cristina Marques de Oliveira, Filipa Trovão, DQ - Departamento de Química, LAQV@REQUIMTE, UCIBIO - Applied Molecular Biosciences Unit, and Universidade do Minho
- Subjects
Models, Molecular ,0301 basic medicine ,Mutant ,Cell ,Crystallography, X-Ray ,Protein Structure, Secondary ,law.invention ,lcsh:Chemistry ,chemistry.chemical_compound ,law ,lcsh:QH301-705.5 ,Spectroscopy ,Anticancer therapy ,mutant p53R280K ,crystal structure ,DNA binding ,anticancer therapy ,General Medicine ,3. Good health ,Cell biology ,Computer Science Applications ,Zinc ,medicine.anatomical_structure ,Recombinant DNA ,Protein Binding ,Programmed cell death ,Arginine ,DNA-binding protein ,Article ,Catalysis ,Inorganic Chemistry ,03 medical and health sciences ,SDG 3 - Good Health and Well-being ,medicine ,Humans ,Physical and Theoretical Chemistry ,Molecular Biology ,Science & Technology ,Mutant p53R280K ,Lysine ,Crystal structure ,Organic Chemistry ,Rational design ,Water ,Hydrogen Bonding ,DNA ,DNA-binding domain ,030104 developmental biology ,chemistry ,lcsh:Biology (General) ,lcsh:QD1-999 ,Mutation ,Tumor Suppressor Protein p53 - Abstract
The p53 tumor suppressor is widely found to be mutated in human cancer. This protein is regarded as a molecular hub regulating different cell responses, namely cell death. Compelling data have demonstrated that the impairment of p53 activity correlates with tumor development and maintenance. For these reasons, the reactivation of p53 function is regarded as a promising strategy to halt cancer. In the present work, the recombinant mutant p53R280K DNA binding domain (DBD) was produced for the first time, and its crystal structure was determined in the absence of DNA to a resolution of 2.0 Å. The solved structure contains four molecules in the asymmetric unit, four zinc(II) ions, and 336 water molecules. The structure was compared with the wild-type p53 DBD structure, isolated and in complex with DNA. These comparisons contributed to a deeper understanding of the mutant p53R280K structure, as well as the loss of DNA binding related to halted transcriptional activity. The structural information derived may also contribute to the rational design of mutant p53 reactivating molecules with potential application in cancer treatment., We thank Gilberto Fronza (from Mutagenesi e Prevenzione Oncologica, Ospedale Policlinico San Martino, Genova, Italy), for providing us with the pLS76 vector. We acknowledge the European Synchrotron Radiation Facility for the provision of synchrotron radiation facilities and access to beamline ID30B. This work received financial support from the European Union (FEDER, Fundo Europeu de Desenvolvimento Regional, funds POCI/01/0145/FEDER/007728 through Programa Operacional Factores de Competitividade–COMPETE) and National Funds (FCT/MCTES, Fundação para a Ciência e Tecnologia and Ministério da Ciência, Tecnologia e Ensino Superior) under the Partnership Agreement PT2020 UID/MULTI/04378/2013, and projects (3599-PPCDT) PTDC/DTP-FTO/1981/2014–POCI-01-0145-FEDER-016581 and RECI/BBB-BEP/0124/2012. FCT fellowships: PD/BD/114046/2015 (Ana Sara Gomes) and SFRH/BD/96189/2013 (Sara Gomes) (thanks FCT PhD Doctoral Programme BiotechHealth), and SFRH/BPD/110640/2015 (Carla Oliveira)., info:eu-repo/semantics/publishedVersion
- Published
- 2018