PXD060074-1

PXD060074 is an original dataset announced via ProteomeXchange.

Title	Reduced structural rigidity of MDMX protein enhances binding to TP53 mRNA
Description	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 the RING domain as a key element, modulated by the presence of other domains. Hydrogen-deuterium exchange mass spectrometry (HDX-MS) and binding assays in high salt conditions demonstrate that the whole protein participates in RNA interaction, with the C-terminal domain likely providing the contact with RNA by electrostatic forces. Modulating pH to influence amino acid protonation showed that the full-length protein binding to RNA is more resilient to pH changes than its truncated C-terminus (322-490). We show that protein structural changes induced by the chelating agent EDTA or the reducing agent TCEP enhances 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.
HostingRepository	PRIDE
AnnounceDate	2026-01-05
AnnouncementXML	Submission_2026-01-04_16:14:49.681.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Lenka Hernychova
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: NEWT:9606;
ModificationList	No PTMs are included in the dataset
Instrument	LTQ Orbitrap Elite

Revision	Datetime	Status	ChangeLog Entry
0	2025-01-22 12:47:08	ID requested
⏵ 1	2026-01-04 16:14:50	announced

10.1042/bsr20253646;

Kucerikova M, Bonczek O, Olivares-Illana V, Rodriguez-Rodriguez A, Sampedro JG, Hernychova L, Hrabal V, Zatloukalova P, Krejcir R, Fahraeus R, Coates PJ, Vojtesek B, Martinkova L, Reduced structural rigidity of MDMX protein enhances binding to TP53 mRNA. Biosci Rep, 45(11):683-96(2025) [pubmed]

submitter keyword: Human, MDMX-MDM2 interaction, p53 mRNA stabilization, RING domain, HDX-MS mapping, LC-MS/MS

Lucia Martinkova
contact affiliation	Masaryk Memorial Cancer Institute, Zluty kopec, Brno, Czech Republic
contact email	lucia.martinkova@mou.cz
lab head
Lenka Hernychova
contact affiliation	Masaryk Memorial Cancer Institutre, Zluty kopec 7, Brno 656 53, Czech Republic
contact email	lenka.hernychova@mou.cz
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD060074
     1. Label: PRIDE project
     2. Name: Reduced structural rigidity of MDMX protein enhances binding to TP53 mRNA