Multisite aggregation of p53 and implications for drug rescue

Wang, GuoZhen; Fersht, Alan R.

doi:10.1073/pnas.1700308114

Cited by 72 publications

(90 citation statements)

References 56 publications

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“…The observed fibrillation delay is likely due to the known slow nucleation of amyloid structures. 13,65 The R248 p53 at 6.5 38 where it may obstruct the migration of the p53 molecules to a fibril nucleus. The accelerated fibril growth in the presence of Ficoll suggests that after nucleation, the fibrils emerge from the clusters and grow in the p53 solution.…”

Section: The Enlarged Aggregation Capacity Of P53 R248qmentioning

confidence: 99%

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Mesoscopic Liquid Clusters Represent a Distinct Condensate of Mutant p53

Yang

Saeedi

Davtyan

et al. 2020

Preprint

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The oncogenic properties of mutant p53 have been ascribed to destabilization of the p53 conformation, followed by aggregation into insoluble fibrils. Here we combine immunofluorescent 3D confocal microscopy of breast cancer cells expressing the p53 mutant Arg248Gln (R248Q) with light scattering from solutions of the purified protein and molecular simulations to probe the mechanisms that govern phase behaviors of the mutant across multiple length scales, from cellular to molecular. We establish that p53 R248Q forms mesoscopic proteinrich clusters, an anomalous liquid phase with several unique properties. We demonstrate that the clusters host and facilitate the nucleation of amyloid fibrils. The distinct characteristics of the clusters of R248Q and wild-type p53 and theoretical models indicate that p53 condensation into clusters is driven by the structural destabilization of the core domain and not by interactions of its extensive disordered region. Two-step nucleation of mutant p53 amyloids suggests means to IntroductionThe wild type p53 protein is a potent cancer suppressor, which is inactivated in almost every tumor, either through mutations in the TP53 gene (in 50% or more of human cancers) or deregulation of its associated pathways. 1-6 By contrast, p53 mutants emerge as potent cancer promoters because they exert a dominant-negative (DN) effect on the wild-type variant and also display oncogenic gain-of-function (GOF) properties by inhibiting other cancer suppressors. 1 Several mechanisms of cancer promotion by mutant p53 have been discussed. [6][7][8][9][10][11][12] It was recently suggested that the structural destabilization of the p53 mutants and the associated aggregation into insoluble, degradation resistant structures, designated fibrils, may play a decisive role in their oncogenicity. 2,[13][14][15] Concurrently, fibril suppression, for instance, by stabilization of the mutant p53 conformation, has been identified as a general way to fight cancer. 2,16 A parallel development identified a new form of biological organization, liquid-liquid phase separation, which constitutes several common membraneless organelles such as nucleoli, Cajal bodies, and P granules. [17][18][19][20][21] Many proteins forming dense liquid condensates in live cells incorporate significant disordered regions and condensation at the low concentrations of these proteins in the nucleoplasm and cytoplasm has been attributed to stronger intermolecular attraction due to the unstructured segments. 19,20 Furthermore, it is well-appreciated that protein dense liquids can serve as precursors and facilitators of higher order organization such as ordering into crystals 22-25 and the formation of polymers of sickle hemoglobin. 26,27 Theoretical analyses and recent experiments suggest that a similar precursor mechanism would apply to the nucleation of amyloid fibrils. [28][29][30][31][32][33]

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Section: The Enlarged Aggregation Capacity Of P53 R248qmentioning

confidence: 99%

“…known to promote p53 aggregation. 14,65 The mutant structures are, however, very different. The largest changes are associated with the N-terminal and C-terminal tails that are both found in various unbound conformations (Figure 7b -d).…”

Section: The Enlarged Aggregation Capacity Of P53 R248qmentioning

confidence: 99%

Mesoscopic Liquid Clusters Represent a Distinct Condensate of Mutant p53

Yang

Saeedi

Davtyan

et al. 2020

Preprint

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“…The altered functions of mutp53 are intertwined with the amyloidogenic process of p53 aggregation. As previously demonstrated by Wang and Fersht (, , ) and Wilcken, Wang, Boeckler, and Fersht (), p53 misfolds or unfolds into an aggregation‐prone stage that loses its DNA‐binding capacity. After the transition stage, p53 rapidly aggregates, by itself or with other proteins, into oligomers and subsequently into fibrils (Figure ), and finally the aggregates accumulate and become resistant to degradation (Wang & Fersht, ).…”

Section: Introductionmentioning

confidence: 55%

Salvation of the fallen angel: Reactivating mutant p53

Yang

Wang

Chen

et al. 2019

British J Pharmacology

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The transcription factor p53 is known as the guardian of the genome for its powerful anti-tumour capacity. However, mutations of p53 that undermine their protein structure, resulting in loss of tumour suppressor function and gain of oncogenic function, have been implicated in more than half of human cancers. The crucial role of mutant forms of p53 in cancer makes it an attractive therapeutic target. A large number of candidates, including low MW compounds, peptides, and nucleic acids, have been identified or designed to rescue p53 mutants and reactivate their anti-tumour capacity through a variety of mechanisms. In this review, we summarize the progress made in the reactivation of mutant forms of p53, focusing on the pharmacological mechanisms of the reactivators of p53 mutants.

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“…The p53C domain contains a single Zn 2+ ion that is required for proper protein folding and function . Mutations to p53C can disrupt protein stability and/or cause loss of Zn . Consequently, the discovery of small molecule Zn chaperones aimed to restore wild‐type function in mutant p53 has generated much attention .…”

Section: Introductionmentioning

confidence: 59%

“…These results demonstrate that an iodinated‐phenol core can be a starting point for envisioning new molecular designs. In addition to protein unfolding, p53 Y220C is prone to the loss of Zn 2+ in the DNA‐binding domain, presenting a new opportunity for drug design.…”

Section: Introductionmentioning

confidence: 99%

Multifunctional Compounds for Activation of the p53‐Y220C Mutant in Cancer

Miller

Orvain

Jozi

et al. 2018

Chemistry A European J

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The p53 protein plays a major role in cancer prevention, and over 50 % of cancer diagnoses can be attributed to p53 malfunction. The common p53 mutation Y220C causes local protein unfolding, aggregation, and can result in a loss of Zn in the DNA-binding domain. Structural analysis has shown that this mutant creates a surface site that can be stabilized using small molecules, and herein a multifunctional approach to restore function to p53-Y220C is reported. A series of compounds has been designed that contain iodinated phenols aimed for interaction and stabilization of the p53-Y220C surface cavity, and Zn-binding fragments for metallochaperone activity. Their Zn-binding affinity was characterized using spectroscopic methods and demonstrate the ability of compounds L4 and L5 to increase intracellular levels of Zn in a p53-Y220C-mutant cell line. The in vitro cytotoxicity of our compounds was initially screened by the National Cancer Institute (NCI-60), followed by testing in three stomach cancer cell lines with varying p53 status', including AGS (WTp53), MKN1 (V143A), and NUGC3 (Y220C). Our most promising ligand, L5, is nearly 3-fold more cytotoxic than cisplatin in a large number of cell lines. The impressive cytotoxicity of L5 is further maintained in a NUGC3 3D spheroid model. L5 also induces Y220C-specific apoptosis in a cleaved caspase-3 assay, reduces levels of unfolded mutant p53, and recovers p53 transcriptional function in the NUGC3 cell line. These results show that these multifunctional scaffolds have the potential to restore wild-type function in mutant p53-Y220C.

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Multisite aggregation of p53 and implications for drug rescue

Cited by 72 publications

References 56 publications

Mesoscopic Liquid Clusters Represent a Distinct Condensate of Mutant p53

Mesoscopic Liquid Clusters Represent a Distinct Condensate of Mutant p53

Salvation of the fallen angel: Reactivating mutant p53

Multifunctional Compounds for Activation of the p53‐Y220C Mutant in Cancer

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