The crystal structure of the R280K mutant of human p53 explains the loss of DNA binding

Ana Sara Gomes, Filipa Trovão, Benedita Andrade Pinheiro, Filipe Freire, Sara Gomes, Carla Oliveira, Lucília Domingues, Maria João Romão, Lucília Saraiva, Ana Luísa Carvalho

Research output: Contribution to journalArticle

3 Citations (Scopus)
6 Downloads (Pure)

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.

Original languageEnglish
Article number1184
JournalInternational Journal of Molecular Sciences
Volume19
Issue number4
DOIs
Publication statusPublished - 13 Apr 2018

Fingerprint

DNA
deoxyribonucleic acid
Crystal structure
crystal structure
cancer
Neoplasms
Molecules
Tumors
tumors
suppressors
molecules
hubs
Oncology
impairment
Cell death
death
maintenance
Zinc
Cell Death
zinc

Keywords

  • Anticancer therapy
  • Crystal structure
  • DNA binding
  • Mutant p53R280K

Cite this

@article{63d940dfe2394ebc96ce67c68661488c,
title = "The crystal structure of the R280K mutant of human p53 explains the loss of DNA binding",
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 {\AA}. 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.",
keywords = "Anticancer therapy, Crystal structure, DNA binding, Mutant p53R280K",
author = "Gomes, {Ana Sara} and Filipa Trov{\~a}o and {Andrade Pinheiro}, Benedita and Filipe Freire and Sara Gomes and Carla Oliveira and Luc{\'i}lia Domingues and Rom{\~a}o, {Maria Jo{\~a}o} and Luc{\'i}lia Saraiva and Carvalho, {Ana Lu{\'i}sa}",
note = "info:eu-repo/grantAgreement/FCT/5876/147258/PT# info:eu-repo/grantAgreement/FCT/3599-PPCDT/125363/PT# PD/BD/114046/2015 SFRH/BD/96189/2013 SFRH/BPD/110640/2015 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, Funda{\cc}{\~a}o para a Ci{\^e}ncia e Tecnologia and Minist{\'e}rio da Ci{\^e}ncia, 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).",
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The crystal structure of the R280K mutant of human p53 explains the loss of DNA binding. / Gomes, Ana Sara; Trovão, Filipa; Andrade Pinheiro, Benedita; Freire, Filipe; Gomes, Sara; Oliveira, Carla; Domingues, Lucília; Romão, Maria João; Saraiva, Lucília; Carvalho, Ana Luísa.

In: International Journal of Molecular Sciences, Vol. 19, No. 4, 1184, 13.04.2018.

Research output: Contribution to journalArticle

TY - JOUR

T1 - The crystal structure of the R280K mutant of human p53 explains the loss of DNA binding

AU - Gomes, Ana Sara

AU - Trovão, Filipa

AU - Andrade Pinheiro, Benedita

AU - Freire, Filipe

AU - Gomes, Sara

AU - Oliveira, Carla

AU - Domingues, Lucília

AU - Romão, Maria João

AU - Saraiva, Lucília

AU - Carvalho, Ana Luísa

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PY - 2018/4/13

Y1 - 2018/4/13

N2 - 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.

AB - 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.

KW - Anticancer therapy

KW - Crystal structure

KW - DNA binding

KW - Mutant p53R280K

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U2 - 10.3390/ijms19041184

DO - 10.3390/ijms19041184

M3 - Article

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JO - International Journal of Molecular Sciences

JF - International Journal of Molecular Sciences

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