Cytoplasmic localization of Mdm2 in cells expressing mutated NPM is mediated by p53
![Thumbnail](/bitstream/handle/20.500.14178/2306/Strachotova-2023-Cytoplasmic%20Localization%20of%20M.pdf.jpg?sequence=5&isAllowed=y)
Autor
Holoubek, Ales
Wolfova, Katerina
Datum vydání
2023Publikováno v
FEBS JournalRočník / Číslo vydání
290 (17)ISBN / ISSN
ISSN: 1742-464XMetadata
Zobrazit celý záznamKolekce
Tato publikace má vydavatelskou verzi s DOI 10.1111/febs.16810
Abstrakt
Specific C-terminal nucleophosmin (NPM) mutations are related to the acute myeloid leukaemia and cause mistargeting of mutated NPM (NPMmut) to the cytoplasm. Consequently, multiple NPM-interacting partners, e.g., the tumour suppressor p53, become also mislocalized. We found that ubiquitin ligase Mdm2 mislocalizes to the cytoplasm in the presence of NPMmut as well. Since p53 interacts with Mdm2, we searched for the NPMmut-p53-Mdm2 complex and interactions of its constituents in live cells and cell lysates using fluorescently tagged proteins, fluorescence lifetime imaging and immunoprecipitation. We proved existence of the ternary complex, which likely adopts a chain-like configuration. Interaction between Mdm2 and NPMmut was not detected, even under conditions of upregulated Mdm2 and p53 induced by Actinomycin D. We assume that p53 serves in the complex as a bridging link between Mdm2 and NPMmut. This conclusion was supported by disruption of the Mdm2-p53 interaction by Nutlin-3A, which resulted in relocalization of Mdm2 to the nucleus, while both NPMmut and p53 remained in the cytoplasm. Importantly, silencing of p53 also prevented mislocalization of Mdm2 in the presence of NPMmut.
Klíčová slova
acute myeloid leukaemia, FRET-FLIM, Mdm2, nucleophosmin mutation, p53
Trvalý odkaz
https://hdl.handle.net/20.500.14178/2306Licence
Licence pro užití plného textu výsledku: Creative Commons Uveďte původ 4.0 International