Mapping of R-SNARE function at distinct intracellular GLUT4 trafficking steps in adipocytes

Dumaine Williams, Jeffrey E. Pessin

Research output: Contribution to journalArticle

72 Scopus citations

Abstract

The functional trafficking steps used by soluble NSF attachment protein receptor (SNARE) proteins have been difficult to establish because of substantial overlap in subcellular localization and because in vitro SNARE-dependent binding and fusion reactions can be promiscuous. Therefore, to functionally identify the site of action of the vesicle-associated membrane protein (VAMP) family of R-SNAREs, we have taken advantage of the temporal requirements of adipocyte biosynthetic sorting of a dual-tagged GLUT4 reporter (myc-GLUT4-GFP) coupled with small interfering RNA gene silencing. Using this approach, we confirm the requirement of VAMP2 and VAMP7 for insulin and osmotic shock trafficking from the vesicle storage sites, respectively, and fusion with the plasma membrane. Moreover, we identify a requirement for VAMP4 for the initial biosynthetic entry of GLUT4 from the Golgi apparatus into the insulin-responsive vesicle compartment, VAMP8, for plasma membrane endocytosis and VAMP2 for sorting to the specialized insulin-responsive compartment after plasma membrane endocytosis.

Original languageEnglish (US)
Pages (from-to)375-387
Number of pages13
JournalJournal of Cell Biology
Volume180
Issue number2
DOIs
StatePublished - Jan 28 2008

ASJC Scopus subject areas

  • Cell Biology

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