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PMID:18227281

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Citation

Williams, D and Pessin, JE (2008) Mapping of R-SNARE function at distinct intracellular GLUT4 trafficking steps in adipocytes. J. Cell Biol. 180:375-87

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.

Links

PubMed PMC2213575 Online version:10.1083/jcb.200709108

Keywords

Adipocytes/metabolism; Animals; Cell Line; Endocytosis; Glucose Transporter Type 4/metabolism; Insulin/metabolism; Mice; Osmotic Pressure; Protein Transport; R-SNARE Proteins/genetics; R-SNARE Proteins/metabolism; RNA, Small Interfering

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

MOUSE:VAMP8

acts_upstream_of_or_within

GO:0030100: regulation of endocytosis

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

MOUSE:VAMP4

acts_upstream_of_or_within

GO:0042996: regulation of Golgi to plasma membrane protein transport

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

MOUSE:GLUT4

acts_upstream_of_or_within

GO:0032869: cellular response to insulin stimulus

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

MOUSE:GLUT4

located_in

GO:0032593: insulin-responsive compartment

ECO:0000314: direct assay evidence used in manual assertion

C

  • part_of:(CL:0000136)

Seeded From UniProt

complete

MOUSE:GLUT4

acts_upstream_of_or_within

GO:0071470: cellular response to osmotic stress

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

MOUSE:VAMP2

acts_upstream_of_or_within

GO:0032869: cellular response to insulin stimulus

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

MOUSE:VAMP2

acts_upstream_of_or_within

GO:0060627: regulation of vesicle-mediated transport

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

MOUSE:VAMP2

acts_upstream_of_or_within

GO:0090316: positive regulation of intracellular protein transport

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

MOUSE:VAMP7

acts_upstream_of_or_within

GO:1900483: regulation of protein targeting to vacuolar membrane

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete


See also

References

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