Working Paper Article Version 1 This version is not peer-reviewed

AML-related NPM Mutations Drive p53 Delocalization Into the Cytoplasm With Possible Impact on p53-Dependent Stress Response

Version 1 : Received: 27 May 2021 / Approved: 28 May 2021 / Online: 28 May 2021 (13:35:48 CEST)

How to cite: Holoubek, A.; Strachotová, D.; Otevřelová, P.; Röselová, P.; Heřman, P.; Brodská, B. AML-related NPM Mutations Drive p53 Delocalization Into the Cytoplasm With Possible Impact on p53-Dependent Stress Response. Preprints 2021, 2021050703 Holoubek, A.; Strachotová, D.; Otevřelová, P.; Röselová, P.; Heřman, P.; Brodská, B. AML-related NPM Mutations Drive p53 Delocalization Into the Cytoplasm With Possible Impact on p53-Dependent Stress Response. Preprints 2021, 2021050703

Abstract

NPM interaction with tumor suppressor p53 is a part of a complex interaction network and considerably affects cellular stress response. An impact of characteristic AML-associated NPM mutations on interaction with p53 has not been investigated yet, although consequences of NPMmut-induced p53 export to the cytoplasm are important for understanding of leukemogenic potential of these mutations. We investigated p53-NPM interaction in live HEK-293T cells by FLIM-FRET and in cell lysates by immunoprecipitation. Results were confirmed in leukemia cell lines. eGFP lifetime-photoconversion was used to follow redistribution dynamics of NPMmut and p53 in Selinexor-treated cells. We confirmed the p53-NPMwt interaction in intact cells and newly documented that this interaction is not compromised by the NPM mutation causing displacement of p53 to the cytoplasm. Importantly, the interaction was not abolished for non-oligomerizing NPM variants with truncated oligomerization domain, suggesting that oligomerization is not essential for interaction of NPM forms with p53. Inhibition of the nuclear exporter XPO1 by Selinexor caused expected nuclear relocalization of both NPMmut and p53. However, significantly different return rates of these proteins indicate nontrivial mechanism of p53 and NPMmut cellular trafficking. We suggest that the altered p53 regulation in cells expressing NPMmut offers a new target for AML therapy.

Subject Areas

p53; nucleophosmin; mutation; acute myeloid leukemia; FLIM-FRET; Selinexor; photoconversion

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