EEF1A is a novel component of the mammalian nuclear protein export machinery

Mireille Khacho, Karim Mekhail, Karine Pilon-Larose, Arnim Pause, Jocelyn Côté, Stephen Lee

Research output: Contribution to journalArticle

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Abstract

The cytoplasmic translation factor eEF1A has been implicated in the nuclear export of tRNA species in lower eukaryotes. Here we demonstrate that eEF1A plays a central role in nuclear export of proteins in mammalian cells. TD-NEM (transcription-dependent nuclear export motif), a newly characterized nuclear export signal, mediates efficient nuclear export of several proteins including the von Hippel-Lindau (VHL) tumor suppressor and the poly(A)-binding protein (PABP1) in a manner that is dependent on ongoing RNA polymerase II (RNA PolII)-dependent transcription. eEF1A interacts specifically with TD-NEM of VHL and PABP1 and disrupting this interaction, by point mutations of key TD-NEM residues or treatment with actinomycin D, an inhibitor of RNA PolII-dependent transcription, prevents assembly and nuclear export. siRNA-induced knockdown or antibody-mediated depletion of eEF1A prevents in vivo and in vitro nuclear export of TD-NEM-containing proteins. Nuclear retention experiments and inhibition of the Exportin-5 pathway suggest that eEF1A stimulates nuclear export of proteins from the cytoplasmic side of the nuclear envelope, without entering the nucleus. Together, these data identify a role for eEF1A, a cytoplasmic mediator of tRNA export in yeast, in the nuclear export of proteins in mammalian cells. These results also provide a link between the translational apparatus and subcellular trafficking machinery demonstrating that these two central pathways in basic metabolism can act cooperatively.

Original languageEnglish (US)
Pages (from-to)5296-5308
Number of pages13
JournalMolecular Biology of the Cell
Volume19
Issue number12
DOIs
StatePublished - Dec 2008
Externally publishedYes

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Cell Nucleus Active Transport
Nuclear Proteins
RNA Polymerase II
Transfer RNA
Von Hippel-Lindau Tumor Suppressor Protein
Poly(A)-Binding Proteins
Nuclear Export Signals
Karyopherins
Amino Acid Motifs
Nuclear Envelope
Dactinomycin
Eukaryota
Point Mutation
Small Interfering RNA
Yeasts

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

Cite this

EEF1A is a novel component of the mammalian nuclear protein export machinery. / Khacho, Mireille; Mekhail, Karim; Pilon-Larose, Karine; Pause, Arnim; Côté, Jocelyn; Lee, Stephen.

In: Molecular Biology of the Cell, Vol. 19, No. 12, 12.2008, p. 5296-5308.

Research output: Contribution to journalArticle

Khacho, M, Mekhail, K, Pilon-Larose, K, Pause, A, Côté, J & Lee, S 2008, 'EEF1A is a novel component of the mammalian nuclear protein export machinery', Molecular Biology of the Cell, vol. 19, no. 12, pp. 5296-5308. https://doi.org/10.1091/mbc.E08-06-0562
Khacho, Mireille ; Mekhail, Karim ; Pilon-Larose, Karine ; Pause, Arnim ; Côté, Jocelyn ; Lee, Stephen. / EEF1A is a novel component of the mammalian nuclear protein export machinery. In: Molecular Biology of the Cell. 2008 ; Vol. 19, No. 12. pp. 5296-5308.
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