An essential role for hGle1 nucleocytoplasmic shuttling in mRNA export

Frederic Kendirgi, Dianne M. Barry, Eric R. Griffis, Maureen A. Powers, Susan R. Wente

    Research output: Contribution to journalArticle

    42 Citations (Scopus)

    Abstract

    Gle1 is required for mRNA export in yeast and human cells. Here, we report that two human Gle1 (hGle1) isoforms are expressed in HeLa cells (hGle1A and B). The two encoded proteins are identical except for their COOH-terminal regions. hGle1A ends with a unique four-amino acid segment, whereas hGle1B has a COOH-terminal 43-amino acid span. Only hGle1B, the more abundant isoform, localizes to the nuclear envelope (NE) and pore complex. To test whether hGle1 is a dynamic shuttling transport factor, we microinjected HeLa cells with recombinant hGle1 and conducted photobleaching studies of live HeLa cells expressing EGFP-hGle1. Both strategies show that hGle1 shuttles between the nucleus and cytoplasm. An internal 39-amino acid domain is necessary and sufficient for mediating nucleocytoplasmic transport. Using a cell-permeable peptide strategy, we document a role for hGle1 shuttling in mRNA export. An hGle1 shuttling domain (SD) peptide impairs the export of both total poly(A)+ RNA and the specific dihydrofolate reductase mRNA. Coincidentally, SD peptide-treated cells show decreased endogenous hGle1 localization at the NE and reduced nucleocytoplasmic shuttling of microinjected, recombinant hGle1. These findings pinpoint the first functional motif in hGle1 and link hGle1 to the dynamic mRNA export mechanism.
    Original languageEnglish
    Pages (from-to)1029-40
    Number of pages12
    JournalJournal of Cell Biology
    Volume160
    Issue number7
    DOIs
    Publication statusPublished - 2003

    Fingerprint

    Messenger RNA
    HeLa Cells
    Nuclear Envelope
    Amino Acids
    Protein Isoforms
    Photobleaching
    Nuclear Pore
    Tetrahydrofolate Dehydrogenase
    Cell Nucleus Active Transport
    Cytoplasm
    Yeasts
    Peptides
    Proteins

    Cite this

    Kendirgi, F., Barry, D. M., Griffis, E. R., Powers, M. A., & Wente, S. R. (2003). An essential role for hGle1 nucleocytoplasmic shuttling in mRNA export. Journal of Cell Biology, 160(7), 1029-40. https://doi.org/10.1083/jcb.200211081
    Kendirgi, Frederic ; Barry, Dianne M. ; Griffis, Eric R. ; Powers, Maureen A. ; Wente, Susan R. / An essential role for hGle1 nucleocytoplasmic shuttling in mRNA export. In: Journal of Cell Biology. 2003 ; Vol. 160, No. 7. pp. 1029-40.
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    abstract = "Gle1 is required for mRNA export in yeast and human cells. Here, we report that two human Gle1 (hGle1) isoforms are expressed in HeLa cells (hGle1A and B). The two encoded proteins are identical except for their COOH-terminal regions. hGle1A ends with a unique four-amino acid segment, whereas hGle1B has a COOH-terminal 43-amino acid span. Only hGle1B, the more abundant isoform, localizes to the nuclear envelope (NE) and pore complex. To test whether hGle1 is a dynamic shuttling transport factor, we microinjected HeLa cells with recombinant hGle1 and conducted photobleaching studies of live HeLa cells expressing EGFP-hGle1. Both strategies show that hGle1 shuttles between the nucleus and cytoplasm. An internal 39-amino acid domain is necessary and sufficient for mediating nucleocytoplasmic transport. Using a cell-permeable peptide strategy, we document a role for hGle1 shuttling in mRNA export. An hGle1 shuttling domain (SD) peptide impairs the export of both total poly(A)+ RNA and the specific dihydrofolate reductase mRNA. Coincidentally, SD peptide-treated cells show decreased endogenous hGle1 localization at the NE and reduced nucleocytoplasmic shuttling of microinjected, recombinant hGle1. These findings pinpoint the first functional motif in hGle1 and link hGle1 to the dynamic mRNA export mechanism.",
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    Kendirgi, F, Barry, DM, Griffis, ER, Powers, MA & Wente, SR 2003, 'An essential role for hGle1 nucleocytoplasmic shuttling in mRNA export', Journal of Cell Biology, vol. 160, no. 7, pp. 1029-40. https://doi.org/10.1083/jcb.200211081

    An essential role for hGle1 nucleocytoplasmic shuttling in mRNA export. / Kendirgi, Frederic; Barry, Dianne M.; Griffis, Eric R.; Powers, Maureen A.; Wente, Susan R.

    In: Journal of Cell Biology, Vol. 160, No. 7, 2003, p. 1029-40.

    Research output: Contribution to journalArticle

    TY - JOUR

    T1 - An essential role for hGle1 nucleocytoplasmic shuttling in mRNA export

    AU - Kendirgi, Frederic

    AU - Barry, Dianne M.

    AU - Griffis, Eric R.

    AU - Powers, Maureen A.

    AU - Wente, Susan R.

    PY - 2003

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    N2 - Gle1 is required for mRNA export in yeast and human cells. Here, we report that two human Gle1 (hGle1) isoforms are expressed in HeLa cells (hGle1A and B). The two encoded proteins are identical except for their COOH-terminal regions. hGle1A ends with a unique four-amino acid segment, whereas hGle1B has a COOH-terminal 43-amino acid span. Only hGle1B, the more abundant isoform, localizes to the nuclear envelope (NE) and pore complex. To test whether hGle1 is a dynamic shuttling transport factor, we microinjected HeLa cells with recombinant hGle1 and conducted photobleaching studies of live HeLa cells expressing EGFP-hGle1. Both strategies show that hGle1 shuttles between the nucleus and cytoplasm. An internal 39-amino acid domain is necessary and sufficient for mediating nucleocytoplasmic transport. Using a cell-permeable peptide strategy, we document a role for hGle1 shuttling in mRNA export. An hGle1 shuttling domain (SD) peptide impairs the export of both total poly(A)+ RNA and the specific dihydrofolate reductase mRNA. Coincidentally, SD peptide-treated cells show decreased endogenous hGle1 localization at the NE and reduced nucleocytoplasmic shuttling of microinjected, recombinant hGle1. These findings pinpoint the first functional motif in hGle1 and link hGle1 to the dynamic mRNA export mechanism.

    AB - Gle1 is required for mRNA export in yeast and human cells. Here, we report that two human Gle1 (hGle1) isoforms are expressed in HeLa cells (hGle1A and B). The two encoded proteins are identical except for their COOH-terminal regions. hGle1A ends with a unique four-amino acid segment, whereas hGle1B has a COOH-terminal 43-amino acid span. Only hGle1B, the more abundant isoform, localizes to the nuclear envelope (NE) and pore complex. To test whether hGle1 is a dynamic shuttling transport factor, we microinjected HeLa cells with recombinant hGle1 and conducted photobleaching studies of live HeLa cells expressing EGFP-hGle1. Both strategies show that hGle1 shuttles between the nucleus and cytoplasm. An internal 39-amino acid domain is necessary and sufficient for mediating nucleocytoplasmic transport. Using a cell-permeable peptide strategy, we document a role for hGle1 shuttling in mRNA export. An hGle1 shuttling domain (SD) peptide impairs the export of both total poly(A)+ RNA and the specific dihydrofolate reductase mRNA. Coincidentally, SD peptide-treated cells show decreased endogenous hGle1 localization at the NE and reduced nucleocytoplasmic shuttling of microinjected, recombinant hGle1. These findings pinpoint the first functional motif in hGle1 and link hGle1 to the dynamic mRNA export mechanism.

    U2 - 10.1083/jcb.200211081

    DO - 10.1083/jcb.200211081

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    JO - Journal of Cell Biology

    JF - Journal of Cell Biology

    SN - 0021-9525

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    ER -