Membrane insertion and intracellular transport of yeast syntaxin Sso2p in mammalian cells

Jussi Jäntti, Sirkka Keränen, Jaana Toikkanen, Esa Kuismanen, Christian Ehnholm, Hans Söderlund, Vesa Olkkonen

    Research output: Contribution to journalArticleScientificpeer-review

    48 Citations (Scopus)

    Abstract

    Proteins of the syntaxin family are suggested to play a key role in determining the specificity of intracellular membrane fusion events. They belong to the class of membrane proteins which are devoid of N-terminal signal sequence and have a C-terminal membrane anchor. Sso2p is a syntaxin homologue involved in the Golgi to plasma membrane vesicular transport in yeast. The protein was transiently expressed in BHK-21 cells using the Semliki Forest virus vector, and its localization and mode of membrane insertion were studied. By immunofluorescence and immuno-EM we show that Sso2p is transported to its final location, the plasma membrane, along the biosynthetic pathway. Experiments with synchronized Sso2p synthesis or expression of the protein in the presence of brefeldin A indicate endoplasmic reticulum as the initial membrane insertion site. During a 20 degrees C temperature block Sso2p accumulated in the Golgi complex and was chased to the plasma membrane by a subsequent 37 degrees C incubation in the presence of cycloheximide. The in vitro translated protein was able to associate with dog pancreatic microsomes post-translationally. A truncated form of Sso2p lacking the putative membrane anchor was used to show that this sequence is necessary for the membrane insertion in vivo and in vitro. The results show that this syntaxin-like protein does not directly associate with its target membrane but uses the secretory pathway to reach its cellular location, raising interesting questions concerning regulation of SNARE-type protein function.
    Original languageEnglish
    Pages (from-to)3623-3633
    Number of pages11
    JournalJournal of Cell Science
    Volume107
    Issue number12
    Publication statusPublished - 1994
    MoE publication typeA1 Journal article-refereed

    Fingerprint

    Dive into the research topics of 'Membrane insertion and intracellular transport of yeast syntaxin Sso2p in mammalian cells'. Together they form a unique fingerprint.

    Cite this