The Paralogue of the Intrinsically Disordered Nuclear Protein 1 Has a Nuclear Localization Sequence that Binds to Human Importin alpha 3

Numerous carrier proteins intervene in protein transport from the cytoplasm to the nucleus in eukaryotic cells. One of those is importin alpha, with several human isoforms; among them, importin alpha 3 (Imp alpha 3) features a particularly high flexibility. The protein NUPR1L is an intrinsically dis...

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Autores: Neira, JL, Rizzuti, B, Jimenez-Alesanco, A, Abian, O, Velazquez-Campoy, A, Iovanna, JL
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2020
País:España
Institución:Fundación para el Fomento de la Investigación Sanitaria y Biomédica de la Comunitat Valenciana (FISABIO)
Repositorio:r-FISABIO. Repositorio Institucional de Producción Científica
OAI Identifier:oai:fisabio.fundanetsuite.com:p12804
Acceso en línea:https://fisabio.portalinvestigacion.com/publicaciones/12804
Access Level:acceso abierto
Palabra clave:circular dichroism
fluorescence
importin
intrinsically disordered protein (IDP)
isothermal titration calorimetry (ITC)
molecular docking
nuclear magnetic resonance (NMR)
paralogue
peptide
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spelling The Paralogue of the Intrinsically Disordered Nuclear Protein 1 Has a Nuclear Localization Sequence that Binds to Human Importin alpha 3Neira, JLRizzuti, BJimenez-Alesanco, AAbian, OVelazquez-Campoy, AIovanna, JLcircular dichroismfluorescenceimportinintrinsically disordered protein (IDP)isothermal titration calorimetry (ITC)molecular dockingnuclear magnetic resonance (NMR)paraloguepeptideNumerous carrier proteins intervene in protein transport from the cytoplasm to the nucleus in eukaryotic cells. One of those is importin alpha, with several human isoforms; among them, importin alpha 3 (Imp alpha 3) 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 Imp alpha 3 and its depleted species, increment Imp alpha 3, 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 alpha 3 with a low micromolar affinity (similar to 5 mu M). 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 increment Imp alpha 3 showed a larger affinity (i.e., lower dissociation constant) compared with that of Imp alpha 3, confirming that the IBB could act as an auto-inhibition region of Imp alpha 3. 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.MDPI2020info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttps://fisabio.portalinvestigacion.com/publicaciones/12804INTERNATIONAL JOURNAL OF MOLECULAR SCIENCESISSN: 16616596ISSNe: 14220067reponame:r-FISABIO. Repositorio Institucional de Producción Científicainstname:Fundación para el Fomento de la Investigación Sanitaria y Biomédica de la Comunitat Valenciana (FISABIO)Inglésinfo:eu-repo/semantics/openAccessoai:fisabio.fundanetsuite.com:p128042026-06-11T12:45:17Z
dc.title.none.fl_str_mv The Paralogue of the Intrinsically Disordered Nuclear Protein 1 Has a Nuclear Localization Sequence that Binds to Human Importin alpha 3
title The Paralogue of the Intrinsically Disordered Nuclear Protein 1 Has a Nuclear Localization Sequence that Binds to Human Importin alpha 3
spellingShingle The Paralogue of the Intrinsically Disordered Nuclear Protein 1 Has a Nuclear Localization Sequence that Binds to Human Importin alpha 3
Neira, JL
circular dichroism
fluorescence
importin
intrinsically disordered protein (IDP)
isothermal titration calorimetry (ITC)
molecular docking
nuclear magnetic resonance (NMR)
paralogue
peptide
title_short The Paralogue of the Intrinsically Disordered Nuclear Protein 1 Has a Nuclear Localization Sequence that Binds to Human Importin alpha 3
title_full The Paralogue of the Intrinsically Disordered Nuclear Protein 1 Has a Nuclear Localization Sequence that Binds to Human Importin alpha 3
title_fullStr The Paralogue of the Intrinsically Disordered Nuclear Protein 1 Has a Nuclear Localization Sequence that Binds to Human Importin alpha 3
title_full_unstemmed The Paralogue of the Intrinsically Disordered Nuclear Protein 1 Has a Nuclear Localization Sequence that Binds to Human Importin alpha 3
title_sort The Paralogue of the Intrinsically Disordered Nuclear Protein 1 Has a Nuclear Localization Sequence that Binds to Human Importin alpha 3
dc.creator.none.fl_str_mv Neira, JL
Rizzuti, B
Jimenez-Alesanco, A
Abian, O
Velazquez-Campoy, A
Iovanna, JL
author Neira, JL
author_facet Neira, JL
Rizzuti, B
Jimenez-Alesanco, A
Abian, O
Velazquez-Campoy, A
Iovanna, JL
author_role author
author2 Rizzuti, B
Jimenez-Alesanco, A
Abian, O
Velazquez-Campoy, A
Iovanna, JL
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv circular dichroism
fluorescence
importin
intrinsically disordered protein (IDP)
isothermal titration calorimetry (ITC)
molecular docking
nuclear magnetic resonance (NMR)
paralogue
peptide
topic circular dichroism
fluorescence
importin
intrinsically disordered protein (IDP)
isothermal titration calorimetry (ITC)
molecular docking
nuclear magnetic resonance (NMR)
paralogue
peptide
description Numerous carrier proteins intervene in protein transport from the cytoplasm to the nucleus in eukaryotic cells. One of those is importin alpha, with several human isoforms; among them, importin alpha 3 (Imp alpha 3) 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 Imp alpha 3 and its depleted species, increment Imp alpha 3, 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 alpha 3 with a low micromolar affinity (similar to 5 mu M). 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 increment Imp alpha 3 showed a larger affinity (i.e., lower dissociation constant) compared with that of Imp alpha 3, confirming that the IBB could act as an auto-inhibition region of Imp alpha 3. 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.
publishDate 2020
dc.date.none.fl_str_mv 2020
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv https://fisabio.portalinvestigacion.com/publicaciones/12804
url https://fisabio.portalinvestigacion.com/publicaciones/12804
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv MDPI
publisher.none.fl_str_mv MDPI
dc.source.none.fl_str_mv INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
ISSN: 16616596
ISSNe: 14220067
reponame:r-FISABIO. Repositorio Institucional de Producción Científica
instname:Fundación para el Fomento de la Investigación Sanitaria y Biomédica de la Comunitat Valenciana (FISABIO)
instname_str Fundación para el Fomento de la Investigación Sanitaria y Biomédica de la Comunitat Valenciana (FISABIO)
reponame_str r-FISABIO. Repositorio Institucional de Producción Científica
collection r-FISABIO. Repositorio Institucional de Producción Científica
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15,81155

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