Citrullinating enzyme PADI4 and transcriptional repressor RING1B bind in cancer cells
Araujo-Abad, Salome ; Rizzuti, Bruno ; Soto-Conde, Lourdes ; Vidal, Miguel ; Abian, Olga (Universidad de Zaragoza) ; Velazquez-Campoy, Adrian (Universidad de Zaragoza) ; Neira, José L. ; de Juan Romero, Camino
Resumen: Polycomb groups (PcGs) are transcriptional repressors, formed by a complex of several proteins, involved in multicellular development and cancer epigenetics. One of these proteins is the E3 ubiquitin-protein ligase RING1 (or RING1B), associated with the regulation of transcriptional repression and responsible for monoubiquitylation of the histone H2A. On the other hand, PADI4 is one of the human isoforms of a family of enzymes implicated in the conversion of arginine to citrulline, and it is also involved in the development of glioblastoma, among other types of cancers. In this work, we showed the association of PADI4 and RING1B in the nucleus and cytosol in several cancer cell lines by using immunofluorescence and proximity ligation assays. Furthermore, we demonstrated that binding was hampered in the presence of GSK484, an enzymatic PADI4 inhibitor, suggesting that RING1B could bind to the active site of PADI4, as confirmed by protein-protein docking simulations. In vitro and in silico findings showed that binding to PADI4 occurred for the isolated fragments corresponding to both the N-terminal (residues 1–221) and C-terminal (residues 228–336) regions of RING1B. Binding to PADI4 was also hampered by GSK484, as shown by isothermal titration calorimetry (ITC) experiments for the sole N-terminal region, and by both NMR and ITC for the C-terminal one. The dissociation constants between PADI4 and any of the two isolated RING1B fragments were in the low micromolar range (~2–10 μM), as measured by fluorescence and ITC. The interaction between RING1B and PADI4 might imply citrullination of the former, leading to several biological consequences, as well as being of potential therapeutic relevance for improving cancer treatment with the generation of new antigens.
Idioma: Inglés
DOI: 10.1016/j.ijbiomac.2024.133163
Año: 2024
Publicado en: International journal of biological macromolecules 274, Part 1 (2024), 133163 [14 pp.]
ISSN: 0141-8130
Financiación: info:eu-repo/grantAgreement/ES/DGA/E45-23R
Financiación: info:eu-repo/grantAgreement/ES/ISCIII-ERDF-ESF/PI21-00394
Financiación: info:eu-repo/grantAgreement/ES/MICINN/AEI/PID2021-127296OB-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 (2024-07-19-18:29:21)
Visitas y descargas
Idioma: Inglés
DOI: 10.1016/j.ijbiomac.2024.133163
Año: 2024
Publicado en: International journal of biological macromolecules 274, Part 1 (2024), 133163 [14 pp.]
ISSN: 0141-8130
Financiación: info:eu-repo/grantAgreement/ES/DGA/E45-23R
Financiación: info:eu-repo/grantAgreement/ES/ISCIII-ERDF-ESF/PI21-00394
Financiación: info:eu-repo/grantAgreement/ES/MICINN/AEI/PID2021-127296OB-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 (2024-07-19-18:29:21)
Permalink:
Visitas y descargas
Este artículo se encuentra en las siguientes colecciones:
Articles
Record created 2024-07-19, last modified 2024-07-19