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Diabetologia
Published in: Diabetologia 8/2015

Open Access 01-08-2015 | Article

Insulin regulates Rab3–Noc2 complex dissociation to promote GLUT4 translocation in rat adipocytes

Authors: Francoise Koumanov, Vinit J. Pereira, Judith D. Richardson, Samantha L. Sargent, Daniel J. Fazakerley, Geoffrey D. Holman

Published in: Diabetologia | Issue 8/2015

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Abstract

Aims/hypothesis

The glucose transporter GLUT4 is present mainly in insulin-responsive tissues of fat, heart and skeletal muscle and is translocated from intracellular membrane compartments to the plasma membrane (PM) upon insulin stimulation. The transit of GLUT4 to the PM is known to be dependent on a series of Rab proteins. However, the extent to which the activity of these Rabs is regulated by the action of insulin action is still unknown. We sought to identify insulin-activated Rab proteins and Rab effectors that facilitate GLUT4 translocation.

Methods

We developed a new photoaffinity reagent (Bio-ATB-GTP) that allows GTP-binding proteomes to be explored. Using this approach we screened for insulin-responsive GTP loading of Rabs in primary rat adipocytes.

Results

We identified Rab3B as a new candidate insulin-stimulated G-protein in adipocytes. Using constitutively active and dominant negative mutants and Rab3 knockdown we provide evidence that Rab3 isoforms are key regulators of GLUT4 translocation in adipocytes. Insulin-stimulated Rab3 GTP binding is associated with disruption of the interaction between Rab3 and its negative effector Noc2. Disruption of the Rab3–Noc2 complex leads to displacement of Noc2 from the PM. This relieves the inhibitory effect of Noc2, facilitating GLUT4 translocation.

Conclusions/interpretation

The discovery of the involvement of Rab3 and Noc2 in an insulin-regulated step in GLUT4 translocation suggests that the control of this translocation process is unexpectedly similar to regulated secretion and particularly pancreatic insulin-vesicle release.
Appendix
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Literature
1.
Stenmark H (2009) Rab GTPases as coordinators of vesicle traffic. Nat Rev Mol Cell Biol 10:513–525PubMedCrossRef
2.
3.
4.
Chen Y, Wang Y, Zhang J et al (2012) Rab10 and myosin-Va mediate insulin-stimulated GLUT4 storage vesicle translocation in adipocytes. J Cell Biol 198:545–560PubMedCentralPubMedCrossRef
5.
Chadt A, Immisch A, de Wendt C et al (2015) Deletion of both Rab-GTPase-activating proteins TBC1D1 and TBC1D4 in mice eliminates insulin- and AICAR-stimulated glucose transport. Diabetes 64:746–759PubMedCrossRef
6.
Tsuboi T, Fukuda M (2006) Rab3A and Rab27A cooperatively regulate the docking step of dense-core vesicle exocytosis in PC12 cells. J Cell Sci 119:2196–2203PubMedCrossRef
7.
Sano H, Eguez L, Teruel MN et al (2007) Rab10, a target of the AS160 Rab GAP, is required for insulin-stimulated translocation of GLUT4 to the adipocyte plasma membrane. Cell Metab 5:293–303PubMedCrossRef
8.
Sun Y, Bilan PJ, Liu Z, Klip A (2010) Rab8A and Rab13 are activated by insulin and regulate GLUT4 translocation in muscle cells. Proc Natl Acad Sci U S A 107:19909–19914PubMedCentralPubMedCrossRef
9.
10.
Taylor LP, Holman GD (1981) Symmetrical kinetic parameters for 3-O-methyl-D-glucose transport in adipocytes in the presence and in the absence of insulin. Biochim Biophys Acta 642:325–335PubMedCrossRef
11.
AlHasani H, Kinck CS, Cushman SW (1998) Endocytosis of the glucose transporter GLUT4 is mediated by the GTPase dynamin. J Biol Chem 273:17504–17510CrossRef
12.
Koumanov F, Pereira VJ, Whitley PR, Holman GD (2012) GLUT4 traffic through an ESCRT-III-dependent sorting compartment in adipocytes. PLoS One 7, e44141PubMedCentralPubMedCrossRef
13.
Simpson IA, Yver DR, Hissin PJ et al (1983) Insulin-stimulated translocation of glucose transporters in the isolated rat adipose cells. Characterization of subcellular fractions. Biochim Biophys Acta 763:393–407PubMedCrossRef
14.
Koumanov F, Richardson JD, Murrow BA, Holman GD (2011) AS160 phosphotyrosine-binding domain constructs inhibit insulin-stimulated GLUT4 vesicle fusion with the plasma membrane. J Biol Chem 286:16574–16582PubMedCentralPubMedCrossRef
15.
16.
Hashimoto M, Hatanaka Y, Yang J, Dhesi J, Holman GD (2001) Synthesis of biotinylated bis(D-glucose) derivatives for glucose transporter photoaffinity labelling. Carbohydr Res 331:119–127PubMedCrossRef
17.
Schwenk RW, Eckel J (2007) A novel method to monitor insulin-stimulated GTP-loading of Rab11a in cardiomyocytes. Cell Signal 19:825–830PubMedCrossRef
18.
Chiang SH, Baumann CA, Kanzaki M et al (2001) Insulin-stimulated GLUT4 translocation requires the CAP-dependent activation of TC10. Nature 410:944–948PubMedCrossRef
19.
Chen XW, Leto D, Chiang SH, Wang Q, Saltiel AR (2007) Activation of RalA is required for insulin-stimulated Glut4 trafficking to the plasma membrane via the exocyst and the motor protein Myo1c. Dev Cell 13:391–404PubMedCrossRef
20.
21.
Guerre-Millo M, Baldini G, Lodish HF, Lavau M, Cushman SW (1997) Rab 3D in rat adipose cells and its overexpression in genetic obesity (Zucker fatty rat). Biochem J 321:89–93PubMedCentralPubMed
22.
Baldini G, Scherer PE, Lodish HF (1995) Nonneuronal expression of Rab3A: induction during adipogenesis and association with different intracellular membranes than Rab3D. Proc Natl Acad Sci U S A 92:4284–4288PubMedCentralPubMedCrossRef
23.
Jewell JL, Oh E, Thurmond DC (2010) Exocytosis mechanisms underlying insulin release and glucose uptake: conserved roles for Munc18c and syntaxin 4. Am J Physiol 298:R517–R531
24.
Cheviet S, Coppola T, Haynes LP, Burgoyne RD, Regazzi R (2004) The Rab-binding protein Noc2 is associated with insulin-containing secretory granules and is essential for pancreatic beta-cell exocytosis. Mol Endocrinol 18:117–126PubMedCrossRef
25.
Schluter OM, Khvotchev M, Jahn R, Sudhof TC (2002) Localization versus function of Rab3 proteins. Evidence for a common regulatory role in controlling fusion. J Biol Chem 277:40919–40929PubMedCrossRef
26.
Piper Hanley K, Hearn T, Berry A et al (2010) In vitro expression of NGN3 identifies RAB3B as the predominant Ras-associated GTP-binding protein 3 family member in human islets. J Endocrinol 207:151–161PubMedCentralPubMedCrossRef
27.
Calderhead DM, Kitagawa K, Tanner LI, Holman GD, Lienhard GE (1990) Insulin regulation of the two glucose transporters in 3T3-L1 adipocytes. J Biol Chem 265:13800–13808
28.
Yang J, Holman GD (1993) Comparison of GLUT4 and GLUT1 subcellular trafficking in basal and insulin-stimulated 3T3-L1 cells. J Biol Chem 268:4600–4603PubMed
29.
30.
Satoh S, Nishimura H, Clark AE et al (1993) Use of bismannose photolabel to elucidate insulin-regulated GLUT4 subcellular trafficking kinetics in rat adipose cells. Evidence that exocytosis is a critical site of hormone action. J Biol Chem 268:17820–17829PubMed
31.
Koumanov F, Jin B, Yang J, Holman GD (2005) Insulin signaling meets vesicle traffic of GLUT4 at a plasma-membrane-activated fusion step. Cell Metab 2:179–189PubMedCrossRef
32.
Lizunov VA, Matsumoto H, Zimmerberg J, Cushman SW, Frolov VA (2005) Insulin stimulates the halting, tethering, and fusion of mobile GLUT4 vesicles in rat adipose cells. J Cell Biol 169:481–489PubMedCentralPubMedCrossRef
33.
Bai L, Wang Y, Fan J et al (2007) Dissecting multiple steps of GLUT4 trafficking and identifying the sites of insulin action. Cell Metab 5:47–57PubMedCrossRef
34.
Cheviet S, Waselle L, Regazzi R (2004) Noc-king out exocrine and endocrine secretion. Trends Cell Biol 14:525–528PubMedCrossRef
35.
Haynes LP, Evans GJ, Morgan A, Burgoyne RD (2001) A direct inhibitory role for the Rab3-specific effector, Noc2, in Ca2+-regulated exocytosis in neuroendocrine cells. J Biol Chem 276:9726–9732PubMedCrossRef
36.
Fukuda N, Emoto M, Nakamori Y et al (2009) DOC2B: a novel syntaxin-4 binding protein mediating insulin-regulated GLUT4 vesicle fusion in adipocytes. Diabetes 58:377–384PubMedCentralPubMedCrossRef
37.
Ramalingam L, Oh E, Yoder SM et al (2012) Doc2b is a key effector of insulin secretion and skeletal muscle insulin sensitivity. Diabetes 61:2424–2432PubMedCentralPubMedCrossRef
38.
39.
Burgoyne RD, Barclay JW, Ciufo LF, Graham ME, Handley MT, Morgan A (2009) The functions of Munc18-1 in regulated exocytosis. Ann N Y Acad Sci 1152:76–86PubMedCrossRef
40.
Gandasi NR, Barg S (2014) Contact-induced clustering of syntaxin and munc18 docks secretory granules at the exocytosis site. Nat Commun 5:3914PubMedCrossRef
41.
Coppola T, Frantz C, Perret-Menoud V, Gattesco S, Hirling H, Regazzi R (2002) Pancreatic beta-cell protein granuphilin binds Rab3 and Munc-18 and controls exocytosis. Mol Biol Cell 13:1906–1915PubMedCentralPubMedCrossRef
42.
Graham ME, Handley MT, Barclay JW et al (2008) A gain-of-function mutant of Munc18-1 stimulates secretory granule recruitment and exocytosis and reveals a direct interaction of Munc18-1 with Rab3. Biochem J 409:407–416PubMedCrossRef
Metadata
Title
Insulin regulates Rab3–Noc2 complex dissociation to promote GLUT4 translocation in rat adipocytes
Authors
Francoise Koumanov
Vinit J. Pereira
Judith D. Richardson
Samantha L. Sargent
Daniel J. Fazakerley
Geoffrey D. Holman
Publication date
01-08-2015
Publisher
Springer Berlin Heidelberg
Published in
Diabetologia / Issue 8/2015
Print ISSN: 0012-186X
Electronic ISSN: 1432-0428
DOI
https://doi.org/10.1007/s00125-015-3627-3

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