Cargando…

Small molecule induced reactivation of mutant p53 in cancer cells

The p53 cancer mutant Y220C is an excellent paradigm for rescuing the function of conformationally unstable p53 mutants because it has a unique surface crevice that can be targeted by small-molecule stabilizers. Here, we have identified a compound, PK7088, which is active in vitro: PK7088 bound to t...

Descripción completa

Detalles Bibliográficos
Autores principales: Liu, Xiangrui, Wilcken, Rainer, Joerger, Andreas C., Chuckowree, Irina S., Amin, Jahangir, Spencer, John, Fersht, Alan R.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3695503/
https://www.ncbi.nlm.nih.gov/pubmed/23630318
http://dx.doi.org/10.1093/nar/gkt305
_version_ 1782274981198036992
author Liu, Xiangrui
Wilcken, Rainer
Joerger, Andreas C.
Chuckowree, Irina S.
Amin, Jahangir
Spencer, John
Fersht, Alan R.
author_facet Liu, Xiangrui
Wilcken, Rainer
Joerger, Andreas C.
Chuckowree, Irina S.
Amin, Jahangir
Spencer, John
Fersht, Alan R.
author_sort Liu, Xiangrui
collection PubMed
description The p53 cancer mutant Y220C is an excellent paradigm for rescuing the function of conformationally unstable p53 mutants because it has a unique surface crevice that can be targeted by small-molecule stabilizers. Here, we have identified a compound, PK7088, which is active in vitro: PK7088 bound to the mutant with a dissociation constant of 140 μM and raised its melting temperature, and we have determined the binding mode of a close structural analogue by X-ray crystallography. We showed that PK7088 is biologically active in cancer cells carrying the Y220C mutant by a battery of tests. PK7088 increased the amount of folded mutant protein with wild-type conformation, as monitored by immunofluorescence, and restored its transcriptional functions. It induced p53-Y220C-dependent growth inhibition, cell-cycle arrest and apoptosis. Most notably, PK7088 increased the expression levels of p21 and the proapoptotic NOXA protein. PK7088 worked synergistically with Nutlin-3 on up-regulating p21 expression, whereas Nutlin-3 on its own had no effect, consistent with its mechanism of action. PK7088 also restored non-transcriptional apoptotic functions of p53 by triggering nuclear export of BAX to the mitochondria. We suggest a set of criteria for assigning activation of p53.
format Online
Article
Text
id pubmed-3695503
institution National Center for Biotechnology Information
language English
publishDate 2013
publisher Oxford University Press
record_format MEDLINE/PubMed
spelling pubmed-36955032013-06-28 Small molecule induced reactivation of mutant p53 in cancer cells Liu, Xiangrui Wilcken, Rainer Joerger, Andreas C. Chuckowree, Irina S. Amin, Jahangir Spencer, John Fersht, Alan R. Nucleic Acids Res Gene Regulation, Chromatin and Epigenetics The p53 cancer mutant Y220C is an excellent paradigm for rescuing the function of conformationally unstable p53 mutants because it has a unique surface crevice that can be targeted by small-molecule stabilizers. Here, we have identified a compound, PK7088, which is active in vitro: PK7088 bound to the mutant with a dissociation constant of 140 μM and raised its melting temperature, and we have determined the binding mode of a close structural analogue by X-ray crystallography. We showed that PK7088 is biologically active in cancer cells carrying the Y220C mutant by a battery of tests. PK7088 increased the amount of folded mutant protein with wild-type conformation, as monitored by immunofluorescence, and restored its transcriptional functions. It induced p53-Y220C-dependent growth inhibition, cell-cycle arrest and apoptosis. Most notably, PK7088 increased the expression levels of p21 and the proapoptotic NOXA protein. PK7088 worked synergistically with Nutlin-3 on up-regulating p21 expression, whereas Nutlin-3 on its own had no effect, consistent with its mechanism of action. PK7088 also restored non-transcriptional apoptotic functions of p53 by triggering nuclear export of BAX to the mitochondria. We suggest a set of criteria for assigning activation of p53. Oxford University Press 2013-07 2013-04-27 /pmc/articles/PMC3695503/ /pubmed/23630318 http://dx.doi.org/10.1093/nar/gkt305 Text en © The Author(s) 2013. Published by Oxford University Press. http://creativecommons.org/licenses/by/3.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Gene Regulation, Chromatin and Epigenetics
Liu, Xiangrui
Wilcken, Rainer
Joerger, Andreas C.
Chuckowree, Irina S.
Amin, Jahangir
Spencer, John
Fersht, Alan R.
Small molecule induced reactivation of mutant p53 in cancer cells
title Small molecule induced reactivation of mutant p53 in cancer cells
title_full Small molecule induced reactivation of mutant p53 in cancer cells
title_fullStr Small molecule induced reactivation of mutant p53 in cancer cells
title_full_unstemmed Small molecule induced reactivation of mutant p53 in cancer cells
title_short Small molecule induced reactivation of mutant p53 in cancer cells
title_sort small molecule induced reactivation of mutant p53 in cancer cells
topic Gene Regulation, Chromatin and Epigenetics
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3695503/
https://www.ncbi.nlm.nih.gov/pubmed/23630318
http://dx.doi.org/10.1093/nar/gkt305
work_keys_str_mv AT liuxiangrui smallmoleculeinducedreactivationofmutantp53incancercells
AT wilckenrainer smallmoleculeinducedreactivationofmutantp53incancercells
AT joergerandreasc smallmoleculeinducedreactivationofmutantp53incancercells
AT chuckowreeirinas smallmoleculeinducedreactivationofmutantp53incancercells
AT aminjahangir smallmoleculeinducedreactivationofmutantp53incancercells
AT spencerjohn smallmoleculeinducedreactivationofmutantp53incancercells
AT fershtalanr smallmoleculeinducedreactivationofmutantp53incancercells