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Cell membrane penetrating function of the nuclear localization sequence in human cytokine IL-1α

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Abstract

Cytokines are released from the cell, bind to their receptors, and affect cellular responses. The precursor form of interleukin 1 alpha (pIL-1α) has a nuclear localization sequence (NLS) that causes it to be localized to the nucleus and regulate specific gene expression. The amino acids of the NLS are basic amino acid-rich sequences, as is the cell penetrating peptide (CPP), which has been widely studied as a way to deliver macromolecules into cells. Here, we hypothesized that the NLS in pIL-1α (pIL-1αNLS) can penetrate the cell membrane and it could deliver macromolecules such as protein in vivo. We characterized cell membrane penetration ability of pIL-1αNLS or its tandem repeated form (2pIL-1αNLS) to enhance its intracellular delivery efficiency. 2pIL-1αNLS showed comparable protein delivery efficiency to TAT–CPP and it mediates endocytosis following heparan sulfate interaction. 2pIL-1αNLS conjugated enhanced green fluorescence protein was localized to the nucleus and the cytoplasm. Intra-peritoneal administration of 2pIL-1αNLS conjugated dTomato protein showed remarkable in vivo intracellular delivery efficiency in various tissues including spleen, liver, and intestine in mice. Moreover, cytotoxicity of 2pIL-1αNLS was not observed even at 100 μM. Our results demonstrate cell membrane-penetrating function of NLS in pIL-1α, which can be used as a safe therapeutic macromolecular delivery peptide.

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Abbreviations

NLS:

Nuclear localization sequence

CPP:

Cell penetrating peptide

IL-1α:

Interleukin 1 alpha

EGFP:

Enhanced green fluorescent protein

MβCD:

Methyl-beta-cyclodextrin

NF-κB:

Nuclear factor kappa-light-chain-enhancer of activated B cells

AP-1:

Activator protein 1

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Acknowledgments

This study is supported by the Basic Science Research Program through the Grants of National Research Foundation of Korea (NRF-2013R1A1A2A10060048) and Hanyang University grant (HY-2011-00000001004).

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Authors and Affiliations

  1. Department of Life Science, Research Institute for Natural Sciences, Hanyang University, Seoul, 133-791, Republic of Korea

    Ja-Hyun Koo, Heeseok Yoon, Won-Ju Kim, Sangho Lim, Hong-Jai Park & Je-Min Choi

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  1. Ja-Hyun Koo
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  2. Heeseok Yoon
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Correspondence to Je-Min Choi.

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Koo, JH., Yoon, H., Kim, WJ. et al. Cell membrane penetrating function of the nuclear localization sequence in human cytokine IL-1α. Mol Biol Rep 41, 8117–8126 (2014). https://doi.org/10.1007/s11033-014-3711-7

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