Syntaxin 6 and Vti1b form a novel SNARE complex, which is up-regulated in activated macrophages to facilitate exocytosis of tumor necrosis Factor-alpha

Rachael Z Murray, Fiona G Wylie, Tatiana Khromykh, David A Hume, Jennifer L Stow

Research output: Contribution to journalArticlepeer-review

Abstract / Description of output

A key function of activated macrophages is to secrete proinflammatory cytokines such as TNFalpha; however, the intracellular pathway and machinery responsible for cytokine trafficking and secretion is largely undefined. Here we show that individual SNARE proteins involved in vesicle docking and fusion are regulated at both gene and protein expression upon stimulation with the bacterial cell wall component lipopolysaccharide. Focusing on two intracellular SNARE proteins, Vti1b and syntaxin 6 (Stx6), we show that they are up-regulated in conjunction with increasing cytokine secretion in activated macrophages and that their levels are selectively titrated to accommodate the volume and timing of post-Golgi cytokine trafficking. In macrophages, Vti1b and syntaxin 6 are localized on intracellular membranes and are present on isolated Golgi membranes and on Golgi-derived TNFalpha vesicles budded in vitro. By immunoprecipitation, we find that Vti1b and syntaxin 6 interact to form a novel intracellular Q-SNARE complex. Functional studies using overexpression of full-length and truncated proteins show that both Vti1b and syntaxin 6 function and have rate-limiting roles in TNFalpha trafficking and secretion. This study shows how macrophages have uniquely adapted a novel Golgi-associated SNARE complex to accommodate their requirement for increased cytokine secretion.
Original languageEnglish
Pages (from-to)10478-83
Number of pages6
JournalJournal of Biological Chemistry
Volume280
Issue number11
DOIs
Publication statusPublished - 18 Mar 2005

Keywords / Materials (for Non-textual outputs)

  • Animals
  • Carrier Proteins
  • Cell Line
  • Cytokines
  • Exocytosis
  • Golgi Apparatus
  • Green Fluorescent Proteins
  • Immunoblotting
  • Immunoprecipitation
  • Inflammation
  • Lipopolysaccharides
  • Macrophages
  • Membrane Proteins
  • Mice
  • Microscopy, Fluorescence
  • Oligonucleotide Array Sequence Analysis
  • Qa-SNARE Proteins
  • Qb-SNARE Proteins
  • RNA
  • SNARE Proteins
  • Time Factors
  • Tumor Necrosis Factor-alpha
  • Up-Regulation
  • Vesicular Transport Proteins

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