Intrinsically disordered chromatin protein NUPR1 binds to the enzyme PADI4

Araujo-Abad, S. ; Neira, J. L. ; Rizzuti, B. ; García-Morales, P. ; Juan Romero, C. de ; Santofimia-Castaño, P. ; Iovanna, J.

Resumen: The 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.

Idioma: Inglés

DOI: 10.1016/j.jmb.2023.168033

Año: 2023

Publicado en: JOURNAL OF MOLECULAR BIOLOGY 435, 8 (2023), 168033 [18 pp.]
ISSN: 0022-2836

Factor impacto SCIMAGO: 2.212 - Biophysics (Q1) - Structural Biology (Q1) - Molecular Biology (Q1)

Financiación: info:eu-repo/grantAgreement/ES/ISCIII/PI22-00824
Financiación: info:eu-repo/grantAgreement/ES/MINECO/CP19-00095

Tipo y forma: Artículo (Versión definitiva)