Next Article in Journal
Circulating miR-99a-5p Expression in Plasma: A Potential Biomarker for Early Diagnosis of Breast Cancer
Next Article in Special Issue
Effects of Detergent on α-Synuclein Structure: A Native MS-Ion Mobility Study
Previous Article in Journal
Discovery of Novel Fetal Hemoglobin Inducers through Small Chemical Library Screening
Previous Article in Special Issue
Relevance of Electrostatic Charges in Compactness, Aggregation, and Phase Separation of Intrinsically Disordered Proteins
Article

The Paralogue of the Intrinsically Disordered Nuclear Protein 1 Has a Nuclear Localization Sequence that Binds to Human Importin α3

1
IDIBE, Universidad Miguel Hernández, 03202 Elche (Alicante), Spain
2
Instituto de Biocomputación y Física de Sistemas Complejos, Joint Units IQFR-CSIC-BIFI, and GBsC-CSIC-BIFI, Universidad de Zaragoza, 50009 Zaragoza, Spain
3
CNR-NANOTEC, Licryl-UOS Cosenza and CEMIF.Cal, Department of Physics, University of Calabria, Via P. Bucci, Cubo 31 C, Arcavacata di Rende, 87036 Cosenza, Italy
4
Instituto de Investigación Sanitaria Aragón (IIS Aragón), 50009 Zaragoza, Spain
5
Centro de Investigación Biomédica en Red en el Área Temática de Enfermedades Hepáticas y Digestivas (CIBERehd), 28029 Madrid, Spain
6
Departamento de Bioquímica y Biología Molecular y Celular, Universidad de Zaragoza, 50009 Zaragoza, Spain
7
Instituto Aragonés de Ciencias de la Salud (IACS), 50009 Zaragoza, Spain
8
Fundacion ARAID, Gobierno de Aragon, 50009 Zaragoza, Spain
9
Centre de Recherche en Cancérologie de Marseille (CRCM), INSERM U1068, CNRS UMR 7258, Aix-Marseille Université and Institut Paoli-Calmettes, Parc Scientifique et Technologique de Luminy, 163 Avenue de Luminy, 13288 Marseille, France
*
Authors to whom correspondence should be addressed.
Int. J. Mol. Sci. 2020, 21(19), 7428; https://0-doi-org.brum.beds.ac.uk/10.3390/ijms21197428
Received: 6 September 2020 / Revised: 28 September 2020 / Accepted: 3 October 2020 / Published: 8 October 2020
Numerous carrier proteins intervene in protein transport from the cytoplasm to the nucleus in eukaryotic cells. One of those is importin α, with several human isoforms; among them, importin α3 (Impα3) 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 Impα3 and its depleted species, ∆Impα3, 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 α3 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 ∆Impα3 showed a larger affinity (i.e., lower dissociation constant) compared with that of Impα3, confirming that the IBB could act as an auto-inhibition region of Impα3. 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. View Full-Text
Keywords: circular dichroism; fluorescence; importin; intrinsically disordered protein (IDP); isothermal titration calorimetry (ITC); molecular docking; nuclear magnetic resonance (NMR); paralogue; peptide circular dichroism; fluorescence; importin; intrinsically disordered protein (IDP); isothermal titration calorimetry (ITC); molecular docking; nuclear magnetic resonance (NMR); paralogue; peptide
Show Figures

Figure 1

MDPI and ACS Style

Neira, J.L.; Rizzuti, B.; Jiménez-Alesanco, A.; Abián, O.; Velázquez-Campoy, A.; Iovanna, J.L. The Paralogue of the Intrinsically Disordered Nuclear Protein 1 Has a Nuclear Localization Sequence that Binds to Human Importin α3. Int. J. Mol. Sci. 2020, 21, 7428. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms21197428

AMA Style

Neira JL, Rizzuti B, Jiménez-Alesanco A, Abián O, Velázquez-Campoy A, Iovanna JL. The Paralogue of the Intrinsically Disordered Nuclear Protein 1 Has a Nuclear Localization Sequence that Binds to Human Importin α3. International Journal of Molecular Sciences. 2020; 21(19):7428. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms21197428

Chicago/Turabian Style

Neira, José L., Bruno Rizzuti, Ana Jiménez-Alesanco, Olga Abián, Adrián Velázquez-Campoy, and Juan L. Iovanna 2020. "The Paralogue of the Intrinsically Disordered Nuclear Protein 1 Has a Nuclear Localization Sequence that Binds to Human Importin α3" International Journal of Molecular Sciences 21, no. 19: 7428. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms21197428

Find Other Styles
Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Article Access Map by Country/Region

1
Back to TopTop