SLMP53-1 interacts with wild-type and mutant p53 DNA-binding domain and reactivates multiple hotspot mutations

Ana Sara Gomes; Helena Ramos; Sara Gomes; Joana B. Loureiro; Joana Soares; Valentina Barcherini; Paola Monti; Gilberto Fronza; Carla Oliveira; Lucília Domingues; Margarida Bastos; Daniel F. A. R. Dourado; Ana Luísa Carvalho; Maria João Romão; Benedita Pinheiro; Filipa Marcelo; Alexandra Carvalho; Maria M. M. Santos; Lucília Saraiva

doi:10.1016/j.bbagen.2019.129440

SLMP53-1 interacts with wild-type and mutant p53 DNA-binding domain and reactivates multiple hotspot mutations

Ana Sara Gomes, Helena Ramos, Sara Gomes, Joana B. Loureiro, Joana Soares, Valentina Barcherini, Paola Monti, Gilberto Fronza, Carla Oliveira, Lucília Domingues, Margarida Bastos, Daniel F. A. R. Dourado, Ana Luísa Carvalho, Maria João Romão, Benedita Pinheiro, Filipa Marcelo, Alexandra Carvalho, Maria M. M. Santos, Lucília Saraiva

MRC PPU

Research output: Contribution to journal › Article › peer-review

15 Citations (Scopus)

Abstract

Background
Half of human cancers harbour TP53 mutations that render p53 inactive as a tumor suppressor. As such, reactivation of mutant (mut)p53 through restoration of wild-type (wt)-like function represents one of the most promising therapeutic strategies in cancer treatment. Recently, we have reported the (S)-tryptophanol-derived oxazoloisoindolinone SLMP53-1 as a new reactivator of wt and mutp53 R280K with in vitro and in vivo p53-dependent antitumor activity. The present work aimed a mechanistic elucidation of mutp53 reactivation by SLMP53-1.

Methods and results
By cellular thermal shift assay (CETSA), it is shown that SLMP53-1 induces wt and mutp53 R280K thermal stabilization, which is indicative of intermolecular interactions with these proteins. Accordingly, in silico studies of wt and mutp53 R280K DNA-binding domain with SLMP53-1 unveiled that the compound binds at the interface of the p53 homodimer with the DNA minor groove. Additionally, using yeast and p53-null tumor cells ectopically expressing distinct highly prevalent mutp53, the ability of SLMP53-1 to reactivate multiple mutp53 is evidenced.

Conclusions
SLMP53-1 is a p53-activating agent with the ability to directly target wt and a set of hotspot mutp53.

General Significance
This work reinforces the encouraging application of SLMP53-1 in the personalized treatment of cancer patients harboring distinct p53 status.

Original language	English
Article number	129440
Pages (from-to)	129440
Journal	BBA - General Subjects
Volume	1864
Issue number	1
Early online date	16 Sept 2019
DOIs	https://doi.org/10.1016/j.bbagen.2019.129440
Publication status	Published - Jan 2020

Keywords

DBD
DNA-binding domain
mutp53
mutant p53
wt
wild-type
CETSA
cellular thermal shift assay
SRB
sulforhodamine B
Reactivator
p53
Chemotherapy
Mutant
Cancer

ASJC Scopus subject areas

Molecular Biology
Biophysics
Biochemistry

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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Cite this

Gomes, A. S., Ramos, H., Gomes, S., Loureiro, J. B., Soares, J., Barcherini, V., Monti, P., Fronza, G., Oliveira, C., Domingues, L., Bastos, M., Dourado, D. F. A. R., Carvalho, A. L., Romão, M. J., Pinheiro, B., Marcelo, F., Carvalho, A., Santos, M. M. M., & Saraiva, L. (2020). SLMP53-1 interacts with wild-type and mutant p53 DNA-binding domain and reactivates multiple hotspot mutations. BBA - General Subjects, 1864(1), 129440. Article 129440. https://doi.org/10.1016/j.bbagen.2019.129440

@article{a69dff1a16e34fa9a62106e094b5daef,

title = "SLMP53-1 interacts with wild-type and mutant p53 DNA-binding domain and reactivates multiple hotspot mutations",

abstract = "BackgroundHalf of human cancers harbour TP53 mutations that render p53 inactive as a tumor suppressor. As such, reactivation of mutant (mut)p53 through restoration of wild-type (wt)-like function represents one of the most promising therapeutic strategies in cancer treatment. Recently, we have reported the (S)-tryptophanol-derived oxazoloisoindolinone SLMP53-1 as a new reactivator of wt and mutp53 R280K with in vitro and in vivo p53-dependent antitumor activity. The present work aimed a mechanistic elucidation of mutp53 reactivation by SLMP53-1.Methods and resultsBy cellular thermal shift assay (CETSA), it is shown that SLMP53-1 induces wt and mutp53 R280K thermal stabilization, which is indicative of intermolecular interactions with these proteins. Accordingly, in silico studies of wt and mutp53 R280K DNA-binding domain with SLMP53-1 unveiled that the compound binds at the interface of the p53 homodimer with the DNA minor groove. Additionally, using yeast and p53-null tumor cells ectopically expressing distinct highly prevalent mutp53, the ability of SLMP53-1 to reactivate multiple mutp53 is evidenced.ConclusionsSLMP53-1 is a p53-activating agent with the ability to directly target wt and a set of hotspot mutp53.General SignificanceThis work reinforces the encouraging application of SLMP53-1 in the personalized treatment of cancer patients harboring distinct p53 status.",

keywords = "DBD, DNA-binding domain, mutp53, mutant p53, wt, wild-type, CETSA, cellular thermal shift assay, SRB, sulforhodamine B, Reactivator, p53, Chemotherapy, Mutant, Cancer",

author = "Gomes, {Ana Sara} and Helena Ramos and Sara Gomes and Loureiro, {Joana B.} and Joana Soares and Valentina Barcherini and Paola Monti and Gilberto Fronza and Carla Oliveira and Luc{\'i}lia Domingues and Margarida Bastos and Dourado, {Daniel F. A. R.} and Carvalho, {Ana Lu{\'i}sa} and Rom{\~a}o, {Maria Jo{\~a}o} and Benedita Pinheiro and Filipa Marcelo and Alexandra Carvalho and Santos, {Maria M. M.} and Luc{\'i}lia Saraiva",

year = "2020",

month = jan,

doi = "10.1016/j.bbagen.2019.129440",

language = "English",

volume = "1864",

pages = "129440",

journal = "BBA - General Subjects",

issn = "0304-4165",

publisher = "Elsevier",

number = "1",

}

Gomes, AS, Ramos, H, Gomes, S, Loureiro, JB, Soares, J, Barcherini, V, Monti, P, Fronza, G, Oliveira, C, Domingues, L, Bastos, M, Dourado, DFAR, Carvalho, AL, Romão, MJ, Pinheiro, B, Marcelo, F, Carvalho, A, Santos, MMM & Saraiva, L 2020, 'SLMP53-1 interacts with wild-type and mutant p53 DNA-binding domain and reactivates multiple hotspot mutations', BBA - General Subjects, vol. 1864, no. 1, 129440, pp. 129440. https://doi.org/10.1016/j.bbagen.2019.129440

TY - JOUR

T1 - SLMP53-1 interacts with wild-type and mutant p53 DNA-binding domain and reactivates multiple hotspot mutations

AU - Gomes, Ana Sara

AU - Ramos, Helena

AU - Gomes, Sara

AU - Loureiro, Joana B.

AU - Soares, Joana

AU - Barcherini, Valentina

AU - Monti, Paola

AU - Fronza, Gilberto

AU - Oliveira, Carla

AU - Domingues, Lucília

AU - Bastos, Margarida

AU - Dourado, Daniel F. A. R.

AU - Carvalho, Ana Luísa

AU - Romão, Maria João

AU - Pinheiro, Benedita

AU - Marcelo, Filipa

AU - Carvalho, Alexandra

AU - Santos, Maria M. M.

AU - Saraiva, Lucília

PY - 2020/1

Y1 - 2020/1

N2 - BackgroundHalf of human cancers harbour TP53 mutations that render p53 inactive as a tumor suppressor. As such, reactivation of mutant (mut)p53 through restoration of wild-type (wt)-like function represents one of the most promising therapeutic strategies in cancer treatment. Recently, we have reported the (S)-tryptophanol-derived oxazoloisoindolinone SLMP53-1 as a new reactivator of wt and mutp53 R280K with in vitro and in vivo p53-dependent antitumor activity. The present work aimed a mechanistic elucidation of mutp53 reactivation by SLMP53-1.Methods and resultsBy cellular thermal shift assay (CETSA), it is shown that SLMP53-1 induces wt and mutp53 R280K thermal stabilization, which is indicative of intermolecular interactions with these proteins. Accordingly, in silico studies of wt and mutp53 R280K DNA-binding domain with SLMP53-1 unveiled that the compound binds at the interface of the p53 homodimer with the DNA minor groove. Additionally, using yeast and p53-null tumor cells ectopically expressing distinct highly prevalent mutp53, the ability of SLMP53-1 to reactivate multiple mutp53 is evidenced.ConclusionsSLMP53-1 is a p53-activating agent with the ability to directly target wt and a set of hotspot mutp53.General SignificanceThis work reinforces the encouraging application of SLMP53-1 in the personalized treatment of cancer patients harboring distinct p53 status.

AB - BackgroundHalf of human cancers harbour TP53 mutations that render p53 inactive as a tumor suppressor. As such, reactivation of mutant (mut)p53 through restoration of wild-type (wt)-like function represents one of the most promising therapeutic strategies in cancer treatment. Recently, we have reported the (S)-tryptophanol-derived oxazoloisoindolinone SLMP53-1 as a new reactivator of wt and mutp53 R280K with in vitro and in vivo p53-dependent antitumor activity. The present work aimed a mechanistic elucidation of mutp53 reactivation by SLMP53-1.Methods and resultsBy cellular thermal shift assay (CETSA), it is shown that SLMP53-1 induces wt and mutp53 R280K thermal stabilization, which is indicative of intermolecular interactions with these proteins. Accordingly, in silico studies of wt and mutp53 R280K DNA-binding domain with SLMP53-1 unveiled that the compound binds at the interface of the p53 homodimer with the DNA minor groove. Additionally, using yeast and p53-null tumor cells ectopically expressing distinct highly prevalent mutp53, the ability of SLMP53-1 to reactivate multiple mutp53 is evidenced.ConclusionsSLMP53-1 is a p53-activating agent with the ability to directly target wt and a set of hotspot mutp53.General SignificanceThis work reinforces the encouraging application of SLMP53-1 in the personalized treatment of cancer patients harboring distinct p53 status.

KW - DBD

KW - DNA-binding domain

KW - mutp53

KW - mutant p53

KW - wt

KW - wild-type

KW - CETSA

KW - cellular thermal shift assay

KW - SRB

KW - sulforhodamine B

KW - Reactivator

KW - p53

KW - Chemotherapy

KW - Mutant

KW - Cancer

UR - http://www.scopus.com/inward/record.url?scp=85073833530&partnerID=8YFLogxK

U2 - 10.1016/j.bbagen.2019.129440

DO - 10.1016/j.bbagen.2019.129440

M3 - Article

C2 - 31536751

SN - 0304-4165

VL - 1864

SP - 129440

JO - BBA - General Subjects

JF - BBA - General Subjects

IS - 1

M1 - 129440

ER -

SLMP53-1 interacts with wild-type and mutant p53 DNA-binding domain and reactivates multiple hotspot mutations

Abstract

Keywords

ASJC Scopus subject areas

UN SDGs

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