000125974 001__ 125974 000125974 005__ 20240705134150.0 000125974 0247_ $$2doi$$a10.1016/j.jmb.2023.168033 000125974 0248_ $$2sideral$$a133523 000125974 037__ $$aART-2023-133523 000125974 041__ $$aeng 000125974 100__ $$aAraujo-Abad, S. 000125974 245__ $$aIntrinsically disordered chromatin protein NUPR1 binds to the enzyme PADI4 000125974 260__ $$c2023 000125974 5060_ $$aAccess copy available to the general public$$fUnrestricted 000125974 5203_ $$aThe nuclear protein 1 (NUPR1) is an intrinsically disordered protein involved in stress-mediated cellular conditions. Its paralogue nuclear protein 1-like (NUPR1L) is p53-regulated, and its expression down-regulates that of the NUPR1 gene. Peptidyl-arginine deiminase 4 (PADI4) is an isoform of a family of enzymes catalyzing arginine to citrulline conversion; it is also involved in stress-mediated cellular conditions. We characterized the interaction between NUPR1 and PADI4 in vitro, in silico, and in cellulo. The interaction of NUPR1 and PADI4 occurred with a dissociation constant of 18 ± 6 μM. The binding region of NUPR1, mapped by NMR, was a hydrophobic polypeptide patch surrounding the key residue Ala33, as pinpointed by: (i) computational results; and, (ii) site-directed mutagenesis of residues of NUPR1. The association between PADI4 and wild-type NUPR1 was also assessed in cellulo by using proximity ligation assays (PLAs) and immunofluorescence (IF), and it occurred mainly in the nucleus. Moreover, binding between NUPR1L and PADI4 also occurred in vitro with an affinity similar to that of NUPR1. Molecular modelling provided information on the binding hot spot for PADI4. This is an example of a disordered partner of PADI4, whereas its other known interacting proteins are well-folded. Altogether, our results suggest that the NUPR1/PADI4 complex could have crucial functions in modulating DNA-repair, favoring metastasis, or facilitating citrullination of other proteins. 000125974 536__ $$9info:eu-repo/grantAgreement/ES/ISCIII/PI22-00824$$9info:eu-repo/grantAgreement/ES/MINECO/CP19-00095 000125974 540__ $$9info:eu-repo/semantics/openAccess$$aby$$uhttp://creativecommons.org/licenses/by/3.0/es/ 000125974 592__ $$a2.212$$b2023 000125974 593__ $$aBiophysics$$c2023$$dQ1 000125974 593__ $$aStructural Biology$$c2023$$dQ1 000125974 593__ $$aMolecular Biology$$c2023$$dQ1 000125974 655_4 $$ainfo:eu-repo/semantics/article$$vinfo:eu-repo/semantics/publishedVersion 000125974 700__ $$aNeira, J. L. 000125974 700__ $$aRizzuti, B. 000125974 700__ $$aGarcía-Morales, P. 000125974 700__ $$aJuan Romero, C. de 000125974 700__ $$aSantofimia-Castaño, P. 000125974 700__ $$aIovanna, J. 000125974 773__ $$g435, 8 (2023), 168033 [18 pp.]$$pJ. Mol. Biol.$$tJOURNAL OF MOLECULAR BIOLOGY$$x0022-2836 000125974 8564_ $$s1425997$$uhttps://zaguan.unizar.es/record/125974/files/texto_completo.pdf$$yVersión publicada 000125974 8564_ $$s2420669$$uhttps://zaguan.unizar.es/record/125974/files/texto_completo.jpg?subformat=icon$$xicon$$yVersión publicada 000125974 909CO $$ooai:zaguan.unizar.es:125974$$particulos$$pdriver 000125974 951__ $$a2024-07-05-12:48:25 000125974 980__ $$aARTICLE