Regulation of nuclear localization signal-importin α interaction by Ca2+/S100A6

Maki Takata, Seiko Shimamoto, Fuminori Yamaguchi, Masaaki Tokuda, Hiroshi Tokumitsu, Ryoji Kobayashi

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Although the precise intracellular roles of S100 proteins are not fully understood, these proteins are thought to be involved in Ca2+-dependent diverse signal transduction pathways. In this report, we identified importin α as a novel target of S100A6. Importin α contains armadillo repeats, essential for binding to nuclear localization signals. Based on the results from GST pull-down assay, gel-shift assay, and co-immunoprecipitation, we demonstrated that S100A6 specifically interacts with the armadillo repeats of importin α in a Ca2+-dependent manner, resulting in inhibition of the nuclear localization signal (NLS)-importin α complex formation in vitro and in vivo. These results indicate S100A6 may regulate the nuclear transport of NLS-cargos in response to increasing concentrations of intracellular Ca2+. Structured summary: MINT- 8045244: Importin alpha (uniprotkb: P52292) physically interacts (MI: 0915) with S100A2 (uniprotkb: P29034) by pull down (MI: 0096). MINT- 8044928: Importin alpha (uniprotkb: P52292) binds (MI: 0407) to S100A6 (uniprotkb: P06703) by pull down (MI: 0096). MINT- 8044941: Importin alpha (uniprotkb: P52292) and S100A6 (uniprotkb: P06703) bind (MI: 0407) by electrophoretic mobility supershift assay (MI: 0412). MINT- 8044997: Importin alpha (uniprotkb: P52292) physically interacts (MI: 0915) with S100A6 (uniprotkb: P06703) by anti bait coimmunoprecipitation (MI: 0006). MINT- 8045031: Importin beta (uniprotkb: Q14974) physically interacts (MI: 0915) with importin alpha (uniprotkb: P52293) and S100A6 (uniprotkb: P06703) by pull down (MI: 0096). MINT- 8044917: Importin alpha (uniprotkb: P52292) binds (MI: 0407) to S100A2 (uniprotkb: P29034) by pull down (MI: 0096). MINT- 8045257: Importin alpha (uniprotkb: P52292) physically interacts (MI: 0915) with S100A6 (uniprotkb: P06703) by pull down (MI: 0096). MINT- 8045015: Importin beta (uniprotkb: Q14974) physically interacts (MI: 0915) with importin alpha (uniprotkb: P52293) and S100A2 (uniprotkb: P29034) by pull down (MI: 0096). MINT- 8045267: Importin alpha (uniprotkb: P52292) physically interacts (MI: 0915) with S100A2 (uniprotkb: P29034) and npm2 (uniprotkb: Q6GQG6) by pull down (MI: 0096). MINT- 8045316: Importin beta (uniprotkb: Q14974) physically interacts (MI: 0915) with importin alpha (uniprotkb: P52293) by pull down (MI: 0096). MINT- 8045302: Importin alpha (uniprotkb: P52292) physically interacts (MI: 0915) with NPM1 (uniprotkb: P06748) and S100A2 (uniprotkb: P29034) by pull down (MI: 0096). MINT- 8045290: Importin alpha (uniprotkb: P52292) physically interacts (MI: 0915) with npm2 (uniprotkb: Q6GQG6) by pull down (MI: 0096). MINT- 8044963, MINT- 8044985: Importin alpha (uniprotkb: P52292) physically interacts (MI: 0915) with S100A2 (uniprotkb: P29034) by anti bait coimmunoprecipitation (MI: 0006). MINT- 8044951: Importin alpha (uniprotkb: P52292) and S100A2 (uniprotkb: P29034) bind (MI: 0407) by electrophoretic mobility supershift assay (MI: 0412).

Original languageEnglish
Pages (from-to)4517-4523
Number of pages7
JournalFEBS Letters
Volume584
Issue number22
DOIs
Publication statusPublished - Nov 19 2010
Externally publishedYes

Fingerprint

alpha Karyopherins
Karyopherins
Nuclear Localization Signals
beta Karyopherins
Assays
Armadillos
Electrophoretic mobility
Electrophoretic Mobility Shift Assay
Signal transduction
Cell Nucleus Active Transport
S100 Proteins

Keywords

  • Armadillo repeat
  • Importin α
  • Nuclear localization signal
  • S100 proteins

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Cell Biology
  • Genetics
  • Molecular Biology
  • Structural Biology

Cite this

Takata, M., Shimamoto, S., Yamaguchi, F., Tokuda, M., Tokumitsu, H., & Kobayashi, R. (2010). Regulation of nuclear localization signal-importin α interaction by Ca2+/S100A6. FEBS Letters, 584(22), 4517-4523. https://doi.org/10.1016/j.febslet.2010.09.052

Regulation of nuclear localization signal-importin α interaction by Ca2+/S100A6. / Takata, Maki; Shimamoto, Seiko; Yamaguchi, Fuminori; Tokuda, Masaaki; Tokumitsu, Hiroshi; Kobayashi, Ryoji.

In: FEBS Letters, Vol. 584, No. 22, 19.11.2010, p. 4517-4523.

Research output: Contribution to journalArticle

Takata, M, Shimamoto, S, Yamaguchi, F, Tokuda, M, Tokumitsu, H & Kobayashi, R 2010, 'Regulation of nuclear localization signal-importin α interaction by Ca2+/S100A6', FEBS Letters, vol. 584, no. 22, pp. 4517-4523. https://doi.org/10.1016/j.febslet.2010.09.052
Takata, Maki ; Shimamoto, Seiko ; Yamaguchi, Fuminori ; Tokuda, Masaaki ; Tokumitsu, Hiroshi ; Kobayashi, Ryoji. / Regulation of nuclear localization signal-importin α interaction by Ca2+/S100A6. In: FEBS Letters. 2010 ; Vol. 584, No. 22. pp. 4517-4523.
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abstract = "Although the precise intracellular roles of S100 proteins are not fully understood, these proteins are thought to be involved in Ca2+-dependent diverse signal transduction pathways. In this report, we identified importin α as a novel target of S100A6. Importin α contains armadillo repeats, essential for binding to nuclear localization signals. Based on the results from GST pull-down assay, gel-shift assay, and co-immunoprecipitation, we demonstrated that S100A6 specifically interacts with the armadillo repeats of importin α in a Ca2+-dependent manner, resulting in inhibition of the nuclear localization signal (NLS)-importin α complex formation in vitro and in vivo. These results indicate S100A6 may regulate the nuclear transport of NLS-cargos in response to increasing concentrations of intracellular Ca2+. Structured summary: MINT- 8045244: Importin alpha (uniprotkb: P52292) physically interacts (MI: 0915) with S100A2 (uniprotkb: P29034) by pull down (MI: 0096). MINT- 8044928: Importin alpha (uniprotkb: P52292) binds (MI: 0407) to S100A6 (uniprotkb: P06703) by pull down (MI: 0096). MINT- 8044941: Importin alpha (uniprotkb: P52292) and S100A6 (uniprotkb: P06703) bind (MI: 0407) by electrophoretic mobility supershift assay (MI: 0412). MINT- 8044997: Importin alpha (uniprotkb: P52292) physically interacts (MI: 0915) with S100A6 (uniprotkb: P06703) by anti bait coimmunoprecipitation (MI: 0006). MINT- 8045031: Importin beta (uniprotkb: Q14974) physically interacts (MI: 0915) with importin alpha (uniprotkb: P52293) and S100A6 (uniprotkb: P06703) by pull down (MI: 0096). MINT- 8044917: Importin alpha (uniprotkb: P52292) binds (MI: 0407) to S100A2 (uniprotkb: P29034) by pull down (MI: 0096). MINT- 8045257: Importin alpha (uniprotkb: P52292) physically interacts (MI: 0915) with S100A6 (uniprotkb: P06703) by pull down (MI: 0096). MINT- 8045015: Importin beta (uniprotkb: Q14974) physically interacts (MI: 0915) with importin alpha (uniprotkb: P52293) and S100A2 (uniprotkb: P29034) by pull down (MI: 0096). MINT- 8045267: Importin alpha (uniprotkb: P52292) physically interacts (MI: 0915) with S100A2 (uniprotkb: P29034) and npm2 (uniprotkb: Q6GQG6) by pull down (MI: 0096). MINT- 8045316: Importin beta (uniprotkb: Q14974) physically interacts (MI: 0915) with importin alpha (uniprotkb: P52293) by pull down (MI: 0096). MINT- 8045302: Importin alpha (uniprotkb: P52292) physically interacts (MI: 0915) with NPM1 (uniprotkb: P06748) and S100A2 (uniprotkb: P29034) by pull down (MI: 0096). MINT- 8045290: Importin alpha (uniprotkb: P52292) physically interacts (MI: 0915) with npm2 (uniprotkb: Q6GQG6) by pull down (MI: 0096). MINT- 8044963, MINT- 8044985: Importin alpha (uniprotkb: P52292) physically interacts (MI: 0915) with S100A2 (uniprotkb: P29034) by anti bait coimmunoprecipitation (MI: 0006). MINT- 8044951: Importin alpha (uniprotkb: P52292) and S100A2 (uniprotkb: P29034) bind (MI: 0407) by electrophoretic mobility supershift assay (MI: 0412).",
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author = "Maki Takata and Seiko Shimamoto and Fuminori Yamaguchi and Masaaki Tokuda and Hiroshi Tokumitsu and Ryoji Kobayashi",
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T1 - Regulation of nuclear localization signal-importin α interaction by Ca2+/S100A6

AU - Takata, Maki

AU - Shimamoto, Seiko

AU - Yamaguchi, Fuminori

AU - Tokuda, Masaaki

AU - Tokumitsu, Hiroshi

AU - Kobayashi, Ryoji

PY - 2010/11/19

Y1 - 2010/11/19

N2 - Although the precise intracellular roles of S100 proteins are not fully understood, these proteins are thought to be involved in Ca2+-dependent diverse signal transduction pathways. In this report, we identified importin α as a novel target of S100A6. Importin α contains armadillo repeats, essential for binding to nuclear localization signals. Based on the results from GST pull-down assay, gel-shift assay, and co-immunoprecipitation, we demonstrated that S100A6 specifically interacts with the armadillo repeats of importin α in a Ca2+-dependent manner, resulting in inhibition of the nuclear localization signal (NLS)-importin α complex formation in vitro and in vivo. These results indicate S100A6 may regulate the nuclear transport of NLS-cargos in response to increasing concentrations of intracellular Ca2+. Structured summary: MINT- 8045244: Importin alpha (uniprotkb: P52292) physically interacts (MI: 0915) with S100A2 (uniprotkb: P29034) by pull down (MI: 0096). MINT- 8044928: Importin alpha (uniprotkb: P52292) binds (MI: 0407) to S100A6 (uniprotkb: P06703) by pull down (MI: 0096). MINT- 8044941: Importin alpha (uniprotkb: P52292) and S100A6 (uniprotkb: P06703) bind (MI: 0407) by electrophoretic mobility supershift assay (MI: 0412). MINT- 8044997: Importin alpha (uniprotkb: P52292) physically interacts (MI: 0915) with S100A6 (uniprotkb: P06703) by anti bait coimmunoprecipitation (MI: 0006). MINT- 8045031: Importin beta (uniprotkb: Q14974) physically interacts (MI: 0915) with importin alpha (uniprotkb: P52293) and S100A6 (uniprotkb: P06703) by pull down (MI: 0096). MINT- 8044917: Importin alpha (uniprotkb: P52292) binds (MI: 0407) to S100A2 (uniprotkb: P29034) by pull down (MI: 0096). MINT- 8045257: Importin alpha (uniprotkb: P52292) physically interacts (MI: 0915) with S100A6 (uniprotkb: P06703) by pull down (MI: 0096). MINT- 8045015: Importin beta (uniprotkb: Q14974) physically interacts (MI: 0915) with importin alpha (uniprotkb: P52293) and S100A2 (uniprotkb: P29034) by pull down (MI: 0096). MINT- 8045267: Importin alpha (uniprotkb: P52292) physically interacts (MI: 0915) with S100A2 (uniprotkb: P29034) and npm2 (uniprotkb: Q6GQG6) by pull down (MI: 0096). MINT- 8045316: Importin beta (uniprotkb: Q14974) physically interacts (MI: 0915) with importin alpha (uniprotkb: P52293) by pull down (MI: 0096). MINT- 8045302: Importin alpha (uniprotkb: P52292) physically interacts (MI: 0915) with NPM1 (uniprotkb: P06748) and S100A2 (uniprotkb: P29034) by pull down (MI: 0096). MINT- 8045290: Importin alpha (uniprotkb: P52292) physically interacts (MI: 0915) with npm2 (uniprotkb: Q6GQG6) by pull down (MI: 0096). MINT- 8044963, MINT- 8044985: Importin alpha (uniprotkb: P52292) physically interacts (MI: 0915) with S100A2 (uniprotkb: P29034) by anti bait coimmunoprecipitation (MI: 0006). MINT- 8044951: Importin alpha (uniprotkb: P52292) and S100A2 (uniprotkb: P29034) bind (MI: 0407) by electrophoretic mobility supershift assay (MI: 0412).

AB - Although the precise intracellular roles of S100 proteins are not fully understood, these proteins are thought to be involved in Ca2+-dependent diverse signal transduction pathways. In this report, we identified importin α as a novel target of S100A6. Importin α contains armadillo repeats, essential for binding to nuclear localization signals. Based on the results from GST pull-down assay, gel-shift assay, and co-immunoprecipitation, we demonstrated that S100A6 specifically interacts with the armadillo repeats of importin α in a Ca2+-dependent manner, resulting in inhibition of the nuclear localization signal (NLS)-importin α complex formation in vitro and in vivo. These results indicate S100A6 may regulate the nuclear transport of NLS-cargos in response to increasing concentrations of intracellular Ca2+. Structured summary: MINT- 8045244: Importin alpha (uniprotkb: P52292) physically interacts (MI: 0915) with S100A2 (uniprotkb: P29034) by pull down (MI: 0096). MINT- 8044928: Importin alpha (uniprotkb: P52292) binds (MI: 0407) to S100A6 (uniprotkb: P06703) by pull down (MI: 0096). MINT- 8044941: Importin alpha (uniprotkb: P52292) and S100A6 (uniprotkb: P06703) bind (MI: 0407) by electrophoretic mobility supershift assay (MI: 0412). MINT- 8044997: Importin alpha (uniprotkb: P52292) physically interacts (MI: 0915) with S100A6 (uniprotkb: P06703) by anti bait coimmunoprecipitation (MI: 0006). MINT- 8045031: Importin beta (uniprotkb: Q14974) physically interacts (MI: 0915) with importin alpha (uniprotkb: P52293) and S100A6 (uniprotkb: P06703) by pull down (MI: 0096). MINT- 8044917: Importin alpha (uniprotkb: P52292) binds (MI: 0407) to S100A2 (uniprotkb: P29034) by pull down (MI: 0096). MINT- 8045257: Importin alpha (uniprotkb: P52292) physically interacts (MI: 0915) with S100A6 (uniprotkb: P06703) by pull down (MI: 0096). MINT- 8045015: Importin beta (uniprotkb: Q14974) physically interacts (MI: 0915) with importin alpha (uniprotkb: P52293) and S100A2 (uniprotkb: P29034) by pull down (MI: 0096). MINT- 8045267: Importin alpha (uniprotkb: P52292) physically interacts (MI: 0915) with S100A2 (uniprotkb: P29034) and npm2 (uniprotkb: Q6GQG6) by pull down (MI: 0096). MINT- 8045316: Importin beta (uniprotkb: Q14974) physically interacts (MI: 0915) with importin alpha (uniprotkb: P52293) by pull down (MI: 0096). MINT- 8045302: Importin alpha (uniprotkb: P52292) physically interacts (MI: 0915) with NPM1 (uniprotkb: P06748) and S100A2 (uniprotkb: P29034) by pull down (MI: 0096). MINT- 8045290: Importin alpha (uniprotkb: P52292) physically interacts (MI: 0915) with npm2 (uniprotkb: Q6GQG6) by pull down (MI: 0096). MINT- 8044963, MINT- 8044985: Importin alpha (uniprotkb: P52292) physically interacts (MI: 0915) with S100A2 (uniprotkb: P29034) by anti bait coimmunoprecipitation (MI: 0006). MINT- 8044951: Importin alpha (uniprotkb: P52292) and S100A2 (uniprotkb: P29034) bind (MI: 0407) by electrophoretic mobility supershift assay (MI: 0412).

KW - Armadillo repeat

KW - Importin α

KW - Nuclear localization signal

KW - S100 proteins

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