Forward and retrograde trafficking in mitotic animal cells ER-Golgi transport arrest restricts protein export from the ER into COPII-coated structures

Theodora Farmaki, Sreenivasan Ponnambalam, Alan R. Prescott, Henrik Clausen, Bor Luen Tang, Wanjin Hong, John M. Lucocq (Lead / Corresponding author)

    Research output: Contribution to journalArticlepeer-review

    61 Citations (Scopus)

    Abstract

    Protein transport arrest occurs between the ER and Golgi stack of mitotic animal cells, but the location of this block is unknown. In this report we use the recycling intermediate compartment protein ERGIC 53/p58 and the plasma membrane protein CD8 to establish the site of transport arrest. Recycled ERGIC 53/p58 and newly synthesised CD8 accumulate in ER cisternae but not in COPII-coated export structures or more distal sites. During mitosis the tubulovesicular ER-related export sites were depleted of the COPII component Sec13p, as shown by immunoelectron microscopy, indicating that COPII budding structures are the target for mitotic inhibition. The extent of recycling of Golgi stack residents was also investigated. In this study we used oligosaccharide modifications on CD8 trapped in the ER of mitotic cells as a sensitive assay for recycling of Golgi stack enzymes. We find that modifications conferred by the Golgi stack-resident GalNac transferase do occur on newly synthesised CD8, but these modifications are entirely due to newly synthesised transferase rather than to enzyme recycled from the Golgi stack. Taken together our findings establish for the first time that the site of ER-Golgi transport arrest of mitotic cells is COPII budding structures, and they clearly speak against a role for recycling in partitioning of Golgi stack proteins via translocation to the ER.

    Original languageEnglish
    Pages (from-to)589-600
    Number of pages12
    JournalJournal of Cell Science
    Volume112
    Issue number5
    Early online date8 Feb 1999
    Publication statusPublished - 1 Mar 1999

    Keywords

    • Budding compartment
    • COPII
    • Golgi
    • Mitosis
    • Recycling

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