000136208 001__ 136208
000136208 005__ 20240719195438.0
000136208 0247_ $$2doi$$a10.1016/j.ijbiomac.2024.133163
000136208 0248_ $$2sideral$$a139080
000136208 037__ $$aART-2024-139080
000136208 041__ $$aeng
000136208 100__ $$aAraujo-Abad, Salome
000136208 245__ $$aCitrullinating enzyme PADI4 and transcriptional repressor RING1B bind in cancer cells
000136208 260__ $$c2024
000136208 5060_ $$aAccess copy available to the general public$$fUnrestricted
000136208 5203_ $$aPolycomb 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.
000136208 536__ $$9info:eu-repo/grantAgreement/ES/DGA/E45-23R$$9info:eu-repo/grantAgreement/ES/ISCIII-ERDF-ESF/PI21-00394$$9info:eu-repo/grantAgreement/ES/MICINN/AEI/PID2021-127296OB-I00
000136208 540__ $$9info:eu-repo/semantics/openAccess$$aby-nc$$uhttp://creativecommons.org/licenses/by-nc/3.0/es/
000136208 655_4 $$ainfo:eu-repo/semantics/article$$vinfo:eu-repo/semantics/publishedVersion
000136208 700__ $$aRizzuti, Bruno
000136208 700__ $$aSoto-Conde, Lourdes
000136208 700__ $$aVidal, Miguel
000136208 700__ $$0(orcid)0000-0001-5664-1729$$aAbian, Olga$$uUniversidad de Zaragoza
000136208 700__ $$0(orcid)0000-0001-5702-4538$$aVelazquez-Campoy, Adrian$$uUniversidad de Zaragoza
000136208 700__ $$aNeira, José L.
000136208 700__ $$ade Juan Romero, Camino
000136208 7102_ $$11002$$2060$$aUniversidad de Zaragoza$$bDpto. Bioq.Biolog.Mol. Celular$$cÁrea Bioquímica y Biolog.Mole.
000136208 773__ $$g274, Part 1 (2024), 133163 [14 pp.]$$pInt. j. biol. macromol.$$tInternational journal of biological macromolecules$$x0141-8130
000136208 8564_ $$s4613603$$uhttps://zaguan.unizar.es/record/136208/files/texto_completo.pdf$$yVersión publicada
000136208 8564_ $$s2700675$$uhttps://zaguan.unizar.es/record/136208/files/texto_completo.jpg?subformat=icon$$xicon$$yVersión publicada
000136208 909CO $$ooai:zaguan.unizar.es:136208$$particulos$$pdriver
000136208 951__ $$a2024-07-19-18:29:21
000136208 980__ $$aARTICLE