96127 20210902121900.0 doi 10.3390/ijms21197428 sideral 120707 ART-2020-120707 eng Neira, J.L. Universidad de Zaragoza (orcid)0000-0003-0668-977X The paralogue of the intrinsically disordered nuclear protein 1 has a nuclear localization sequence that binds to human importin a3 2020 Access copy available to the general public Unrestricted Numerous carrier proteins intervene in protein transport from the cytoplasm to the nucleus in eukaryotic cells. One of those is importin a, with several human isoforms; among them, importin a3 (Impa3) features a particularly high flexibility. The protein NUPR1L is an intrinsically disordered protein (IDP), evolved as a paralogue of nuclear protein 1 (NUPR1), which is involved in chromatin remodeling and DNA repair. It is predicted that NUPR1L has a nuclear localization sequence (NLS) from residues Arg51 to Gln74, in order to allow for nuclear translocation. We studied in this work the ability of intact NUPR1L to bind Impa3 and its depleted species, ¿Impa3, without the importin binding domain (IBB), using fluorescence, isothermal titration calorimetry (ITC), circular dichroism (CD), nuclear magnetic resonance (NMR), and molecular docking techniques. Furthermore, the binding of the peptide matching the isolated NLS region of NUPR1L (NLS-NUPR1L) was also studied using the same methods. Our results show that NUPR1L was bound to Imp a3 with a low micromolar affinity (~5 µM). Furthermore, a similar affinity value was observed for the binding of NLS-NUPR1L. These findings indicate that the NLS region, which was unfolded in isolation in solution, was essentially responsible for the binding of NUPR1L to both importin species. This result was also confirmed by our in silico modeling. The binding reaction of NLS-NUPR1L to ¿Impa3 showed a larger affinity (i.e., lower dissociation constant) compared with that of Impa3, confirming that the IBB could act as an auto-inhibition region of Impa3. Taken together, our findings pinpoint the theoretical predictions of the NLS region in NUPR1L and, more importantly, suggest that this IDP relies on an importin for its nuclear translocation. info:eu-repo/grantAgreement/ES/DGA/B25-17R info:eu-repo/grantAgreement/ES/DGA/E45-17R info:eu-repo/grantAgreement/ES/ISCIII/CIBERehd info:eu-repo/grantAgreement/ES/ISCIII/CPII13-00017 info:eu-repo/grantAgreement/ES/MCIU-AEI-FEDER/BFU2016-78232-P info:eu-repo/grantAgreement/ES/MCIU-AEI-FEDER/RTI2018-097991-B-I00 info:eu-repo/semantics/openAccess by http://creativecommons.org/licenses/by/3.0/es/ 5.923 2020 BIOCHEMISTRY & MOLECULAR BIOLOGY 67 / 297 = 0.226 2020 Q1 T1 CHEMISTRY, MULTIDISCIPLINARY 49 / 178 = 0.275 2020 Q2 T1 1.455 2020 Catalysis 2020 Q1 Computer Science Applications 2020 Q1 Inorganic Chemistry 2020 Q1 Spectroscopy 2020 Q1 Molecular Biology 2020 Q1 Organic Chemistry 2020 Q1 Physical and Theoretical Chemistry 2020 Q1 Medicine (miscellaneous) 2020 Q1 info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion Rizzuti, B. Jiménez-Alesanco, A. Universidad de Zaragoza (orcid)0000-0003-4726-7821 Abián, O. Universidad de Zaragoza (orcid)0000-0001-5664-1729 Velázquez-Campoy, A. Universidad de Zaragoza (orcid)0000-0001-5702-4538 Iovanna, J.L. 1002 060 Universidad de Zaragoza Dpto. Bioq.Biolog.Mol. Celular Área Bioquímica y Biolog.Mole. 5007 570 Universidad de Zaragoza Dpto. Informát.Ingenie.Sistms. Área Lenguajes y Sistemas Inf. 21, 19 (2020), 7428 [19 pp] Int. j. mol. sci. International Journal of Molecular Sciences 1661-6596 754203 http://zaguan.unizar.es/record/96127/files/texto_completo.pdf Versión publicada 509144 http://zaguan.unizar.es/record/96127/files/texto_completo.jpg?subformat=icon icon Versión publicada oai:zaguan.unizar.es:96127 articulos driver 2021-09-02-10:35:03 ARTICLE