Characterization of an importin in α/β-recognized nuclear localization signal in β-dystroglycan

Bárbara Lara-Chacón, Mario Bermúdez De León, Daniel Leocadio, Pablo Gómez, Lizeth Fuentes-Mera, Ivette Martínez-Vieyra, Arturo Ortega, David A. Jans, Bulmaro Cisneros

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

β-dystroglycan (β-DG) is a widely expressed transmembrane protein that plays important roles in connecting the extracellular matrix to the cytoskeleton, and thereby contributing to plasma membrane integrity and signal transduction. We previously observed nuclear localization of β-DG in cultured cell lines, implying the existence of a nuclear targeting mechanism that directs it to the nucleus instead of the plasma membrane. In this study, we delineate the nuclear import pathway of β-DG, characterizing a functional nuclear localization signal (NLS) in the β-DG cytoplasmic domain, within amino acids 776-782. The NLS either alone or in the context of the whole β-DG protein was able to target the heterologous GFP protein to the nucleus, with site-directed mutagenesis indicating that amino acids R779and K780are critical for NLS functionality. The nuclear transport molecules Importin (Imp)α and Impβ bound with high affinity to the NLS of β-DG and were found to be essential for NLS-dependent nuclear import in an in vitro reconstituted nuclear transport assay; cotransfection experiments confirmed the dependence on Ran for nuclear accumulation. Intriguingly, experiments suggested that tyrosine phosphorylation of β-DG may result in cytoplasmic retention, with Y892playing a key role. β-DG thus follows a conventional Impα/β-dependent nuclear import pathway, with important implications for its potential function in the nucleus. © 2010 Wiley-Liss, Inc.
Original languageEnglish
Pages (from-to)706-717
Number of pages12
JournalJournal of Cellular Biochemistry
DOIs
Publication statusPublished - 1 Jun 2010
Externally publishedYes

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Dystroglycans
Karyopherins
Nuclear Localization Signals
Cell Nucleus Active Transport
Cell membranes
Amino Acids
Signal transduction
Mutagenesis
Phosphorylation
Proteins
Tyrosine
Cell Membrane
Assays
Experiments
Cells
Site-Directed Mutagenesis
Molecules
Cytoskeleton
Extracellular Matrix
Cultured Cells

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Lara-Chacón, B., De León, M. B., Leocadio, D., Gómez, P., Fuentes-Mera, L., Martínez-Vieyra, I., ... Cisneros, B. (2010). Characterization of an importin in α/β-recognized nuclear localization signal in β-dystroglycan. Journal of Cellular Biochemistry, 706-717. https://doi.org/10.1002/jcb.22581
Lara-Chacón, Bárbara ; De León, Mario Bermúdez ; Leocadio, Daniel ; Gómez, Pablo ; Fuentes-Mera, Lizeth ; Martínez-Vieyra, Ivette ; Ortega, Arturo ; Jans, David A. ; Cisneros, Bulmaro. / Characterization of an importin in α/β-recognized nuclear localization signal in β-dystroglycan. In: Journal of Cellular Biochemistry. 2010 ; pp. 706-717.
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Lara-Chacón, B, De León, MB, Leocadio, D, Gómez, P, Fuentes-Mera, L, Martínez-Vieyra, I, Ortega, A, Jans, DA & Cisneros, B 2010, 'Characterization of an importin in α/β-recognized nuclear localization signal in β-dystroglycan', Journal of Cellular Biochemistry, pp. 706-717. https://doi.org/10.1002/jcb.22581

Characterization of an importin in α/β-recognized nuclear localization signal in β-dystroglycan. / Lara-Chacón, Bárbara; De León, Mario Bermúdez; Leocadio, Daniel; Gómez, Pablo; Fuentes-Mera, Lizeth; Martínez-Vieyra, Ivette; Ortega, Arturo; Jans, David A.; Cisneros, Bulmaro.

In: Journal of Cellular Biochemistry, 01.06.2010, p. 706-717.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Characterization of an importin in α/β-recognized nuclear localization signal in β-dystroglycan

AU - Lara-Chacón, Bárbara

AU - De León, Mario Bermúdez

AU - Leocadio, Daniel

AU - Gómez, Pablo

AU - Fuentes-Mera, Lizeth

AU - Martínez-Vieyra, Ivette

AU - Ortega, Arturo

AU - Jans, David A.

AU - Cisneros, Bulmaro

PY - 2010/6/1

Y1 - 2010/6/1

N2 - β-dystroglycan (β-DG) is a widely expressed transmembrane protein that plays important roles in connecting the extracellular matrix to the cytoskeleton, and thereby contributing to plasma membrane integrity and signal transduction. We previously observed nuclear localization of β-DG in cultured cell lines, implying the existence of a nuclear targeting mechanism that directs it to the nucleus instead of the plasma membrane. In this study, we delineate the nuclear import pathway of β-DG, characterizing a functional nuclear localization signal (NLS) in the β-DG cytoplasmic domain, within amino acids 776-782. The NLS either alone or in the context of the whole β-DG protein was able to target the heterologous GFP protein to the nucleus, with site-directed mutagenesis indicating that amino acids R779and K780are critical for NLS functionality. The nuclear transport molecules Importin (Imp)α and Impβ bound with high affinity to the NLS of β-DG and were found to be essential for NLS-dependent nuclear import in an in vitro reconstituted nuclear transport assay; cotransfection experiments confirmed the dependence on Ran for nuclear accumulation. Intriguingly, experiments suggested that tyrosine phosphorylation of β-DG may result in cytoplasmic retention, with Y892playing a key role. β-DG thus follows a conventional Impα/β-dependent nuclear import pathway, with important implications for its potential function in the nucleus. © 2010 Wiley-Liss, Inc.

AB - β-dystroglycan (β-DG) is a widely expressed transmembrane protein that plays important roles in connecting the extracellular matrix to the cytoskeleton, and thereby contributing to plasma membrane integrity and signal transduction. We previously observed nuclear localization of β-DG in cultured cell lines, implying the existence of a nuclear targeting mechanism that directs it to the nucleus instead of the plasma membrane. In this study, we delineate the nuclear import pathway of β-DG, characterizing a functional nuclear localization signal (NLS) in the β-DG cytoplasmic domain, within amino acids 776-782. The NLS either alone or in the context of the whole β-DG protein was able to target the heterologous GFP protein to the nucleus, with site-directed mutagenesis indicating that amino acids R779and K780are critical for NLS functionality. The nuclear transport molecules Importin (Imp)α and Impβ bound with high affinity to the NLS of β-DG and were found to be essential for NLS-dependent nuclear import in an in vitro reconstituted nuclear transport assay; cotransfection experiments confirmed the dependence on Ran for nuclear accumulation. Intriguingly, experiments suggested that tyrosine phosphorylation of β-DG may result in cytoplasmic retention, with Y892playing a key role. β-DG thus follows a conventional Impα/β-dependent nuclear import pathway, with important implications for its potential function in the nucleus. © 2010 Wiley-Liss, Inc.

U2 - 10.1002/jcb.22581

DO - 10.1002/jcb.22581

M3 - Article

SP - 706

EP - 717

JO - Journal of Cellular Biochemistry

JF - Journal of Cellular Biochemistry

SN - 0730-2312

ER -