The paralogue of the intrinsically disordered nuclear protein 1 has a nuclear localization sequence that binds to human importin a3

Neira, J.L. (Universidad de Zaragoza) ; Rizzuti, B. ; Jiménez-Alesanco, A. (Universidad de Zaragoza) ; Abián, O. (Universidad de Zaragoza) ; Velázquez-Campoy, A. (Universidad de Zaragoza) ; Iovanna, J.L.

Resumen: 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.

Idioma: Inglés

DOI: 10.3390/ijms21197428

Año: 2020

Publicado en: International Journal of Molecular Sciences 21, 19 (2020), 7428 [19 pp]
ISSN: 1661-6596

Factor impacto JCR: 5.923 (2020)

Categ. JCR: BIOCHEMISTRY & MOLECULAR BIOLOGY rank: 67 / 297 = 0.226 (2020) - Q1 - T1
Categ. JCR: CHEMISTRY, MULTIDISCIPLINARY rank: 49 / 178 = 0.275 (2020) - Q2 - T1

Factor impacto SCIMAGO: 1.455 - Catalysis (Q1) - Computer Science Applications (Q1) - Inorganic Chemistry (Q1) - Spectroscopy (Q1) - Molecular Biology (Q1) - Organic Chemistry (Q1) - Physical and Theoretical Chemistry (Q1) - Medicine (miscellaneous) (Q1)

Financiación: info:eu-repo/grantAgreement/ES/DGA/B25-17R
Financiación: info:eu-repo/grantAgreement/ES/DGA/E45-17R
Financiación: info:eu-repo/grantAgreement/ES/ISCIII/CIBERehd
Financiación: info:eu-repo/grantAgreement/ES/ISCIII/CPII13-00017
Financiación: info:eu-repo/grantAgreement/ES/MCIU-AEI-FEDER/BFU2016-78232-P
Financiación: info:eu-repo/grantAgreement/ES/MCIU-AEI-FEDER/RTI2018-097991-B-I00

Tipo y forma: Article (Published version)

Área (Departamento): Área Bioquímica y Biolog.Mole. (Dpto. Bioq.Biolog.Mol. Celular)
Área (Departamento): Área Lenguajes y Sistemas Inf. (Dpto. Informát.Ingenie.Sistms.)