Syntaxin 6 regulates Glut4 trafficking in 3T3-L1 adipocytes.
Insulin stimulates the movement of glucose transporter-4 (Glut4)-containing vesicles to the plasma membrane of adipose cells. We investigated the role of post-Golgi t-soluble N-ethylmaleimide-sensitive factor attachment protein receptors (SNAREs) in the trafficking of Glut4 in 3T3-L1 adipocytes. Greater than 85% of syntaxin 6 was found in Glut4-containing vesicles, and this t-SNARE exhibited insulin-stimulated movement to the plasma membrane. In contrast, the colocalization of Glut4 with syntaxin 7, 8, or 12/13 was limited and these molecules did not translocate to the plasma membrane. We used adenovirus to overexpress the cytosolic domain of these syntaxin's and studied their effects on Glut4 traffic. Overexpression of the cytosolic domain of syntaxin 6 did not affect insulin-stimulated glucose transport, but increased basal deGlc transport and cell surface Glut4 levels. Moreover, the syntaxin 6 cytosolic domain significantly reduced the rate of Glut4 reinternalization after insulin withdrawal and perturbed subendosomal Glut4 sorting; the corresponding domains of syntaxins 8 and 12 were without effect. Our data suggest that syntaxin 6 is involved in a membrane-trafficking step that sequesters Glut4 away from traffic destined for the plasma membrane. We speculate that this is at the level of traffic of Glut4 into its unique storage compartment and that syntaxin 16 may be involved.
Perera
H Kumudu I
HK
The Henry Wellcome Laboratory of Cell Biology, Division of Biochemistry and Molecular Biology, Institute of Biomedical and Life Sciences, University of Glasgow, Glasgow G12 8QQ, Scotland, United Kingdom.
Clarke
Mairi
M
Morris
Nicholas J
NJ
Hong
Wanjin
W
Chamberlain
Luke H
LH
Gould
Gwyn W
GW
D041721:3T3-L1 Cells; D017667:Adipocytes / Q000378:metabolism* / Q000502:physiology; D000818:Animals; D002451:Cell Compartmentation / Q000502:physiology; D002462:Cell Membrane / Q000502:physiology; D022162:Cytoplasmic Vesicles / Q000502:physiology; D051275:Glucose Transporter Type 4; D007328:Insulin / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism*; D051379:Mice; D009004:Monosaccharide Transport Proteins / Q000378:metabolism* / Q000502:physiology; D009124:Muscle Proteins; D017434:Protein Structure, Tertiary; D021381:Protein Transport / Q000502:physiology; D050765:Qa-SNARE Proteins; D011994:Recombinant Proteins / Q000378:metabolism; D013347:Subcellular Fractions
D051275:Glucose Transporter Type 4; D007328:Insulin; D008565:Membrane Proteins; D009004:Monosaccharide Transport Proteins; D009124:Muscle Proteins; D050765:Qa-SNARE Proteins; D011994:Recombinant Proteins; C495693:Slc2a4 protein, mouse