Multisite phosphorylation of the AML-linked C-terminal of nucleophosmin (NPM1) orchestrates protein stability, DNA binding and charge block-driven phase separation
Rivero-García, Pablo ; Giner-Arroyo, Rafael L ; Tamargo-Azpilicueta, Joaquín ; Telfer, Abbey ; Frezza , Elisa ; Velázquez-Campoy, Adrián (Universidad de Zaragoza) ; Díaz-Moreno, Sofía ; De la Rosa, Miguel A. ; Díaz-Moreno, Irene
Resumen: Nucleophosmin (NPM1) is a nucleolar protein commonly mutated in ~30% of newly diagnosed acute myeloid leukemia (AML) cases. These mutations occur in the terminal exon of the NPM1 gene, affecting the C-terminal DNA-binding domain of the protein and causing its delocalization to the cytoplasm—a hallmark of NPM1-mutated AML. NPM1 shuttling to the nucleoplasm is tightly regulated by posttranslational modifications, such as phosphorylation of Ser254, Ser260, and Tyr271 of the DNA-binding domain. However, the structural mechanisms underlying this process remain unclear. In this work, we show that Ser-to-Asp (S254D–S260D) and Tyr-to-pCMF (para-carboxymethyl phenylalanine) (Y271pCMF) phosphomimetic mutations induce significant structural and dynamical rearrangements, as well as drastic modifications in electrostatic surface potential. These changes compromise recognition of a G-quadruplex sequence from the c-MYC promoter by reducing DNA-binding affinity, reshape histone capturing dynamics, and fade charge segregation in the histone-binding domain. Combination of such substitutions in a triple phosphomimetic variant (S254D–S260D–Y271pCMF) further destabilizes the domain’s structure and triggers protein aggregation. Altogether, these findings suggest that phosphorylation of Ser254, Ser260, and Tyr271 of the C-end DNA-binding domain weakens both DNA affinity and charge block-driven liquid–liquid phase separation, offering a molecular explanation for the delocalization of NPM1 outside of the nucleolus.
Idioma: Inglés
DOI: 10.1093/nar/gkag165
Año: 2026
Publicado en: Nucleic Acids Research 54, 5 (2026), 17 pp.
ISSN: 0305-1048
Financiación: info:eu-repo/grantAgreement/ES/CSIC/JAEINT24-EX-0171
Financiación: info:eu-repo/grantAgreement/ES/MCIU/PID2024-157414NB-I00
Tipo y forma: Article (Published version)
Área (Departamento): Área Bioquímica y Biolog.Mole. (Dpto. Bioq.Biolog.Mol. Celular)
You must give appropriate credit, provide a link to the license, and indicate if changes were made. You may do so in any reasonable manner, but not in any way that suggests the licensor endorses you or your use. You may not use the material for commercial purposes.
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Idioma: Inglés
DOI: 10.1093/nar/gkag165
Año: 2026
Publicado en: Nucleic Acids Research 54, 5 (2026), 17 pp.
ISSN: 0305-1048
Financiación: info:eu-repo/grantAgreement/ES/CSIC/JAEINT24-EX-0171
Financiación: info:eu-repo/grantAgreement/ES/MCIU/PID2024-157414NB-I00
Tipo y forma: Article (Published version)
Área (Departamento): Área Bioquímica y Biolog.Mole. (Dpto. Bioq.Biolog.Mol. Celular)
You must give appropriate credit, provide a link to the license, and indicate if changes were made. You may do so in any reasonable manner, but not in any way that suggests the licensor endorses you or your use. You may not use the material for commercial purposes. Exportado de SIDERAL (2026-03-18-13:52:38)
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Record created 2026-03-18, last modified 2026-03-18