Sec13 shuttles between the nucleus and the cytoplasm and stably interacts with Nup96 at the nuclear pore complex

Jost Enninga, Agata Levay, Beatriz M A Fontoura

Research output: Contribution to journalArticle

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Abstract

Sec13 is a constituent of the endoplasmic reticulum and the nuclear pore complex (NPC). At the endoplasmic reticulum, Sec13 is involved in the biogenesis of COPII-coated vesicles, whereas at the NPC its function is unknown. We show here, by yeast two-hybrid screenings and biochemical assays, that a region at the amino terminus of the human nuclear pore complex protein Nup96 interacts with the WD (Trp-Asp) repeat region of human Sec13. By using immunofluorescence and confocal and immunoelectron microscopy, we found that in interphase, Sec13 and Nup96 are localized at both sides of the NPC in addition to other intracellular sites. In mitosis, Sec13 was found dispersed throughout the cell, whereas a pool of Nup96 colocalized with the spindle apparatus. Photobleaching experiments showed that Sec13 shuttles between intranuclear sites and the cytoplasm, and a fraction of Sec13 is stably associated with NPCs. Cotransfection of Sec13 and the Sec13 binding site of Nup96 decreased the mobile pool of Sec13, demonstrating the interaction of Sec13 and Nup96 in vivo. Targeting studies showed that Sec13 is actively transported into the nucleus and contains a nuclear localization signal. These results indicate that Sec13 stably interacts with Nup96 at the NPC during interphase and that the shuttling of Sec13 between the nucleus and the cytoplasm may couple and regulate functions between these two compartments.

Original languageEnglish (US)
Pages (from-to)7271-7284
Number of pages14
JournalMolecular and Cellular Biology
Volume23
Issue number20
DOIs
StatePublished - Oct 2003

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Nuclear Pore
Cytoplasm
Interphase
Endoplasmic Reticulum
COP-Coated Vesicles
Nuclear Pore Complex Proteins
Photobleaching
Nuclear Localization Signals
Spindle Apparatus
Immunoelectron Microscopy
Fluorescence Microscopy
Mitosis
Confocal Microscopy
nuclear pore complex protein 96
Yeasts
Binding Sites

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cell Biology

Cite this

Sec13 shuttles between the nucleus and the cytoplasm and stably interacts with Nup96 at the nuclear pore complex. / Enninga, Jost; Levay, Agata; Fontoura, Beatriz M A.

In: Molecular and Cellular Biology, Vol. 23, No. 20, 10.2003, p. 7271-7284.

Research output: Contribution to journalArticle

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