Background: Nucleophosmin-1 (NPM1) is an abundant multifunctional protein, implicated in a variety of biological processes and in the pathogenesis of several human malignancies. Its C-terminal domain (CTD) is endowed with a three helix bundle and we demonstrated that several regions within it, associated with acute myeloid leukemia (AML), have a strong tendency to form beta amyloid-like assemblies toxic for cells. The central helix of the bundle (H2) resulted the most amyloidgenic region; here we aim to model the cytoxicity processes of the H2 sequence and getting clues of a potential involvement in toxicity of the interaction between CTDs and cellular membranes. Methods: We investigated the interaction of CTD-NPM1 regions with model membranes through fluorescence, SPR, CD and ESR spectroscopies and the localization of NPM1 by immune-fluorescence in leukemic cells. Results: Our findings indicate that investigated regions are able to interact with membranes with different mechanisms and outlined the importance of the presence of cholesterol. Conclusions: H2 showed a preference of interaction with membrane containing cholesterol determining a sensitive fluidification of the bilayer, while N-term H2 causes a stiffening of central and outer regions of the lipid system. Noticeably, NPM1 mut A demonstrated to thicken at the plasma membrane, differently from wt. These findings were corroborated by diverse mechanisms of interaction of CTDs toward membrane models in vitro. General significance: This study suggests that the direct interaction of several regions of NPM1CTD with cellular membranes could be implicated in diseases where NPM1 is mutated and/or where its overexpression is cytoxic.

Nucleophosmin-1 regions associated with acute myeloid leukemia interact differently with lipid membranes

Federici, Luca;De Cola, Antonella;
2018-01-01

Abstract

Background: Nucleophosmin-1 (NPM1) is an abundant multifunctional protein, implicated in a variety of biological processes and in the pathogenesis of several human malignancies. Its C-terminal domain (CTD) is endowed with a three helix bundle and we demonstrated that several regions within it, associated with acute myeloid leukemia (AML), have a strong tendency to form beta amyloid-like assemblies toxic for cells. The central helix of the bundle (H2) resulted the most amyloidgenic region; here we aim to model the cytoxicity processes of the H2 sequence and getting clues of a potential involvement in toxicity of the interaction between CTDs and cellular membranes. Methods: We investigated the interaction of CTD-NPM1 regions with model membranes through fluorescence, SPR, CD and ESR spectroscopies and the localization of NPM1 by immune-fluorescence in leukemic cells. Results: Our findings indicate that investigated regions are able to interact with membranes with different mechanisms and outlined the importance of the presence of cholesterol. Conclusions: H2 showed a preference of interaction with membrane containing cholesterol determining a sensitive fluidification of the bilayer, while N-term H2 causes a stiffening of central and outer regions of the lipid system. Noticeably, NPM1 mut A demonstrated to thicken at the plasma membrane, differently from wt. These findings were corroborated by diverse mechanisms of interaction of CTDs toward membrane models in vitro. General significance: This study suggests that the direct interaction of several regions of NPM1CTD with cellular membranes could be implicated in diseases where NPM1 is mutated and/or where its overexpression is cytoxic.
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11564/684720
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