Reduced structural rigidity of MDMX protein enhances binding to
TP53 mRNA

J 2025

Reduced structural rigidity of MDMX protein enhances binding to TP53 mRNA

KUČERÍKOVÁ, Martina; Ondrej BONCZEK; Vanesa OLIVARES-ILLANA; Andres RODRIGUEZ-RODRIGUEZ; Jose G. SAMPEDRO et al.

Základní údaje

Originální název

Reduced structural rigidity of MDMX protein enhances binding to TP53 mRNA

Autoři

Vydání

BIOSCIENCE REPORTS, LONDON, PORTLAND PRESS LTD, 2025, 0144-8463

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10608 Biochemistry and molecular biology

Stát vydavatele

Velká Británie a Severní Irsko

Utajení

není předmětem státního či obchodního tajemství

Odkazy

URL

Impakt faktor

Impact factor: 4.700 v roce 2024

Označené pro přenos do RIV

Ano

Kód RIV

RIV/00216224:14310/25:00143440

Organizační jednotka

Přírodovědecká fakulta

Klíčová slova anglicky

TP53mRNA; HDX-MS mapping; MDMX–RNA interaction; RING domain

Štítky

14314010, 143160, rivok

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 30. 1. 2026 10:22, Mgr. Marie Novosadová Šípková, DiS.

Anotace

V originále

The two murine double minute (MDM) family members, MDM2 and MDMX, are a well-established negative regulator of p53 activity. Under DNA damage conditions, MDM2 and MDMX are phosphorylated near their RING domains (serine 395 at MDM2 and serine 403 at MDMX) and switch to act as p53 positive regulators. MDMX binds to TP53 mRNA and acts as a chaperone for RNA structure, enabling MDM2 to bind. This interaction enhances TP53 mRNA translation, leading to increased p53 protein production. While the biological significance of this interaction has been described, the specific features of the MDMX-RNA interaction remain poorly understood. We used various MDMX protein constructs to characterize binding to TP53 mRNA and identified that the interaction mediated by the RING domain is modulated by the presence of other domains. Hydrogen-deuterium exchange mass spectrometry (HDX-MS) and binding assays in high salt conditions and various pH demonstrate that the whole protein participates in RNA interaction, with the C-terminal domain likely providing the contact with RNA by electrostatic forces. We show that protein structural changes induced by the chelating agent EDTA or the reducing agent TCEP enhance RNA binding by promoting partial structural destabilization of the protein. Our findings suggest that the MDMX/TP53 mRNA interaction is complex, with the RING domain binding to RNA and being supported by the entire protein, which acts as a scaffold for the RNA interaction. These results contribute to a better understanding of MDMX's role in TP53 mRNA binding and provide valuable insights for future investigation of the MDM2-MDMX-TP53 mRNA complex, which is crucial for p53 stabilization and activation under DNA-damaging conditions.

Citovat

KUČERÍKOVÁ, Martina; Ondrej BONCZEK; Vanesa OLIVARES-ILLANA; Andres RODRIGUEZ-RODRIGUEZ; Jose G. SAMPEDRO; Lenka HERNYCHOVA; Václav HRABAL; Pavlina ZATLOUKALOVA; Radovan KREJCIR; Robin FAHRAEUS; Philip J. COATES; Bořivoj VOJTĚŠEK a Lucia MARTINKOVÁ. Reduced structural rigidity of MDMX protein enhances binding to TP53 mRNA. BIOSCIENCE REPORTS. LONDON: PORTLAND PRESS LTD, 2025, roč. 45, č. 11, s. 683-696. ISSN 0144-8463. Dostupné z: https://doi.org/10.1042/BSR20253646.

@article{2548403,
   author = {Kučeríková, Martina and Bonczek, Ondrej and OlivaresandIllana, Vanesa and RodriguezandRodriguez, Andres and Sampedro, Jose G. and Hernychova, Lenka and Hrabal, Václav and Zatloukalova, Pavlina and Krejcir, Radovan and Fahraeus, Robin and Coates, Philip J. and Vojtěšek, Bořivoj and Martinková, Lucia},
   article_location = {LONDON},
   article_number = {11},
   doi = {https://doi.org/10.1042/BSR20253646},
   keywords = {TP53mRNA; HDX-MS mapping; MDMX–RNA interaction; RING domain},
   language = {eng},
   issn = {0144-8463},
   journal = {BIOSCIENCE REPORTS},
   title = {Reduced structural rigidity of MDMX protein enhances binding to TP53 mRNA},
   url = {https://portlandpress.com/bioscirep/article/45/11/683/236760/Reduced-structural-rigidity-of-MDMX-protein},
   volume = {45},
   year = {2025}
}

TY  - JOUR
ID  - 2548403
AU  - Kučeríková, Martina - Bonczek, Ondrej - Olivares-Illana, Vanesa - Rodriguez-Rodriguez, Andres - Sampedro, Jose G. - Hernychova, Lenka - Hrabal, Václav - Zatloukalova, Pavlina - Krejcir, Radovan - Fahraeus, Robin - Coates, Philip J. - Vojtěšek, Bořivoj - Martinková, Lucia
PY  - 2025
TI  - Reduced structural rigidity of MDMX protein enhances binding to TP53 mRNA
JF  - BIOSCIENCE REPORTS
VL  - 45
IS  - 11
SP  - 683-696
EP  - 683-696
PB  - PORTLAND PRESS LTD
SN  - 01448463
KW  - TP53mRNA
KW  - HDX-MS mapping
KW  - MDMX–RNA interaction
KW  - RING domain
UR  - https://portlandpress.com/bioscirep/article/45/11/683/236760/Reduced-structural-rigidity-of-MDMX-protein
N2  - The two murine double minute (MDM) family members, MDM2 and MDMX, are a well-established negative regulator of p53 activity. Under DNA damage conditions, MDM2 and MDMX are phosphorylated near their RING domains (serine 395 at MDM2 and serine 403 at MDMX) and switch to act as p53 positive regulators. MDMX binds to TP53 mRNA and acts as a chaperone for RNA structure, enabling MDM2 to bind. This interaction enhances TP53 mRNA translation, leading to increased p53 protein production. While the biological significance of this interaction has been described, the specific features of the MDMX-RNA interaction remain poorly understood. We used various MDMX protein constructs to characterize binding to TP53 mRNA and identified that the interaction mediated by the RING domain is modulated by the presence of other domains. Hydrogen-deuterium exchange mass spectrometry (HDX-MS) and binding assays in high salt conditions and various pH demonstrate that the whole protein participates in RNA interaction, with the C-terminal domain likely providing the contact with RNA by electrostatic forces. We show that protein structural changes induced by the chelating agent EDTA or the reducing agent TCEP enhance RNA binding by promoting partial structural destabilization of the protein. Our findings suggest that the MDMX/TP53 mRNA interaction is complex, with the RING domain binding to RNA and being supported by the entire protein, which acts as a scaffold for the RNA interaction. These results contribute to a better understanding of MDMX's role in TP53 mRNA binding and provide valuable insights for future investigation of the MDM2-MDMX-TP53 mRNA complex, which is crucial for p53 stabilization and activation under DNA-damaging conditions.
ER  -

KUČERÍKOVÁ, Martina; Ondrej BONCZEK; Vanesa OLIVARES-ILLANA; Andres RODRIGUEZ-RODRIGUEZ; Jose G. SAMPEDRO; Lenka HERNYCHOVA; Václav HRABAL; Pavlina ZATLOUKALOVA; Radovan KREJCIR; Robin FAHRAEUS; Philip J. COATES; Bořivoj VOJTĚŠEK a Lucia MARTINKOVÁ. Reduced structural rigidity of MDMX protein enhances binding to TP53 mRNA. \textit{BIOSCIENCE REPORTS}. LONDON: PORTLAND PRESS LTD, 2025, roč.~45, č.~11, s.~683-696. ISSN~0144-8463. Dostupné z: https://doi.org/10.1042/BSR20253646.

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