Molecular identification of 26 syntaxin genes and their assignment to the different trafficking pathways in Paramecium

Roland Kissmehl, Christina Schilde, Thomas Wassmer, Carsten Danzer, Kathrin Nuehse, Kaya Lutter, Helmut Plattner

Research output: Contribution to journalArticle

Abstract

SNARE proteins have been classified as vesicular (v)- and target (t)-SNAREs and play a central role in the various membrane interactions in eukaryotic cells. Based on the Paramecium genome project, we have identified a multigene family of at least 26 members encoding the t-SNARE syntaxin (PtSyx) that can be grouped into 15 subfamilies. Paramecium syntaxins match the classical build-up of syntaxins, being 'tail-anchored' membrane proteins with an N-terminal cytoplasmic domain and a membrane-bound single C-terminal hydrophobic domain. The membrane anchor is preceded by a conserved SNARE domain of approximately 60 amino acids that is supposed to participate in SNARE complex assembly. In a phylogenetic analysis, most of the Paramecium syntaxin genes were found to cluster in groups together with those from other organisms in a pathway-specific manner, allowing an assignment to different compartments in a homology-dependent way. However, some of them seem to have no counterparts in metazoans. In another approach, we fused one representative member of each of the syntaxin isoforms to green fluorescent protein and assessed the in vivo localization, which was further supported by immunolocalization of some syntaxins. This allowed us to assign syntaxins to all important trafficking pathways in Paramecium.
Original languageEnglish
Pages (from-to)523-542
Number of pages20
JournalTraffic
Volume8
Issue number5
DOIs
Publication statusPublished - May 2007

Keywords

  • amino acid sequence
  • vesicular transport proteins
  • biological transport
  • cell membrane
  • endoplasmicreticulum
  • golgi apparatus
  • green fluorescent proteins
  • intracellular membranes
  • immunoelectron microscopy
  • biological models
  • molecular models
  • molecular sequence data
  • paramecium
  • phylogeny
  • protein isoforms
  • tertiary protein structure
  • protozoan proteins
  • Qa-SNARE proteins
  • RNA interference
  • recombinant fusion proteins
  • DNA sequence analysis
  • amino acid sequence homology
  • transfection
  • transport vesicles

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    Kissmehl, R., Schilde, C., Wassmer, T., Danzer, C., Nuehse, K., Lutter, K., & Plattner, H. (2007). Molecular identification of 26 syntaxin genes and their assignment to the different trafficking pathways in Paramecium. Traffic, 8(5), 523-542. https://doi.org/10.1111/j.1600-0854.2007.00544.x