Learn more: PMC Disclaimer | PMC Copyright Notice
Potential role of Rab4 in the regulation of subcellular localization of Glut4 in adipocytes.
Abstract
A role for Rab4 in the translocation of the glucose transporter Glut4 induced by insulin has been recently proposed. To study more directly the role of this small GTPase, freshly isolated adipocytes were transiently transfected with the cDNAs of both an epitope-tagged Glut4-myc and Rab4, a system which allows direct measurement of the concentration of Glut4 molecules at the cell surface. When cells were cotransfected with Glut4-myc and Rab4, the concentration of Glut4-myc at the cell surface decreased in parallel with the increased expression of Rab4, suggesting that Rab4 participates in the intracellular retention of Glut4. In parallel, the amount of Rab4 associated with the Glut4-containing vesicles increased. When Rab4 was moderately overexpressed, the number of Glut4-myc molecules recruited to the cell surface in response to insulin was similar to that observed in mock-transfected cells, and thus the insulin efficiency was increased. When Rab4 was expressed at a higher level, the amount of Glut4-myc present at the cell surface in response to insulin decreased. Since the overexpressed protein was predominantly cytosolic, this suggests that the cytosolic Rab4 might complex some factor(s) necessary for insulin action. This hypothesis was strengthened by the fact that Rab4 deltaCT, a Rab4 mutant lacking the geranylgeranylation sites, inhibited insulin-induced recruitement of Glut4-myc to the cell surface, even when moderately overexpressed. Rab3D was without effect on Glut4-myc subcellular distribution in basal or insulin-stimulated conditions. While two mutated proteins unable to bind GTP did not decrease the number of Glut4-myc molecules in basal or insulin-stimulated conditions at the plasma membrane, the behavior of a mutated Rab4 protein without GTPase activity was similar to that of the wild-type Rab4 protein, indicating that GTP binding but not its hydrolysis was required for the observed effects. Altogether, our results suggest that Rab4, but not Rab3D, participates in the molecular mechanism involved in the subcellular distribution of the Glut4 molecules both in basal and in insulin-stimulated conditions in adipocytes.
Full Text
The Full Text of this article is available as a PDF (353K).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Aledo JC, Darakhshan F, Hundal HS. Rab4, but not the transferrin receptor, is colocalized with GLUT4 in an insulin-sensitive intracellular compartment in rat skeletal muscle. Biochem Biophys Res Commun. 1995 Oct 4;215(1):321–328. [PubMed] [Google Scholar]
- Baldini G, Hohl T, Lin HY, Lodish HF. Cloning of a Rab3 isotype predominantly expressed in adipocytes. Proc Natl Acad Sci U S A. 1992 Jun 1;89(11):5049–5052. [PMC free article] [PubMed] [Google Scholar]
- Baldini G, Hohman R, Charron MJ, Lodish HF. Insulin and nonhydrolyzable GTP analogs induce translocation of GLUT 4 to the plasma membrane in alpha-toxin-permeabilized rat adipose cells. J Biol Chem. 1991 Mar 5;266(7):4037–4040. [PubMed] [Google Scholar]
- Baldini G, Scherer PE, Lodish HF. Nonneuronal expression of Rab3A: induction during adipogenesis and association with different intracellular membranes than Rab3D. Proc Natl Acad Sci U S A. 1995 May 9;92(10):4284–4288. [PMC free article] [PubMed] [Google Scholar]
- Beranger F, Paterson H, Powers S, de Gunzburg J, Hancock JF. The effector domain of Rab6, plus a highly hydrophobic C terminus, is required for Golgi apparatus localization. Mol Cell Biol. 1994 Jan;14(1):744–758. [PMC free article] [PubMed] [Google Scholar]
- Bortoluzzi MN, Cormont M, Gautier N, Van Obberghen E, Le Marchand-Brustel Y. GTPase activating protein activity for Rab4 is enriched in the plasma membrane of 3T3-L1 adipocytes. Possible involvement in the regulation of Rab4 subcellular localization. Diabetologia. 1996 Aug;39(8):899–906. [PubMed] [Google Scholar]
- Brozinick JT, Jr, Yaspelkis BB, 3rd, Wilson CM, Grant KE, Gibbs EM, Cushman SW, Ivy JL. Glucose transport and GLUT4 protein distribution in skeletal muscle of GLUT4 transgenic mice. Biochem J. 1996 Jan 1;313(Pt 1):133–140. [PMC free article] [PubMed] [Google Scholar]
- Cheatham B, Kahn CR. Insulin action and the insulin signaling network. Endocr Rev. 1995 Apr;16(2):117–142. [PubMed] [Google Scholar]
- Chung SH, Takai Y, Holz RW. Evidence that the Rab3a-binding protein, rabphilin3a, enhances regulated secretion. Studies in adrenal chromaffin cells. J Biol Chem. 1995 Jul 14;270(28):16714–16718. [PubMed] [Google Scholar]
- Cormont M, Tanti JF, Grémeaux T, Van Obberghen E, Le Marchand-Brustel Y. Subcellular distribution of low molecular weight guanosine triphosphate-binding proteins in adipocytes: colocalization with the glucose transporter Glut 4. Endocrinology. 1991 Dec;129(6):3343–3350. [PubMed] [Google Scholar]
- Cormont M, Tanti JF, Zahraoui A, Van Obberghen E, Le Marchand-Brustel Y. Rab4 is phosphorylated by the insulin-activated extracellular-signal-regulated kinase ERK1. Eur J Biochem. 1994 Feb 1;219(3):1081–1085. [PubMed] [Google Scholar]
- Cormont M, Tanti JF, Zahraoui A, Van Obberghen E, Tavitian A, Le Marchand-Brustel Y. Insulin and okadaic acid induce Rab4 redistribution in adipocytes. J Biol Chem. 1993 Sep 15;268(26):19491–19497. [PubMed] [Google Scholar]
- Cormont M, Van Obberghen E, Zerial M, Le Marchand-Brustel Y. Insulin induces a change in Rab5 subcellular localization in adipocytes independently of phosphatidylinositol 3-kinase activation. Endocrinology. 1996 Aug;137(8):3408–3415. [PubMed] [Google Scholar]
- Gould GW, Holman GD. The glucose transporter family: structure, function and tissue-specific expression. Biochem J. 1993 Oct 15;295(Pt 2):329–341. [PMC free article] [PubMed] [Google Scholar]
- Hansen PA, Gulve EA, Marshall BA, Gao J, Pessin JE, Holloszy JO, Mueckler M. Skeletal muscle glucose transport and metabolism are enhanced in transgenic mice overexpressing the Glut4 glucose transporter. J Biol Chem. 1995 Jan 27;270(4):1679–1684. [PubMed] [Google Scholar]
- James DE, Piper RC, Slot JW. Insulin stimulation of GLUT-4 translocation: a model for regulated recycling. Trends Cell Biol. 1994 Apr;4(4):120–126. [PubMed] [Google Scholar]
- Kinsella BT, Maltese WA. rab GTP-binding proteins with three different carboxyl-terminal cysteine motifs are modified in vivo by 20-carbon isoprenoids. J Biol Chem. 1992 Feb 25;267(6):3940–3945. [PubMed] [Google Scholar]
- Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. [PubMed] [Google Scholar]
- Le Marchand-Brustel Y, Gautier N, Cormont M, Van Obberghen E. Wortmannin inhibits the action of insulin but not that of okadaic acid in skeletal muscle: comparison with fat cells. Endocrinology. 1995 Aug;136(8):3564–3570. [PubMed] [Google Scholar]
- Le Marchand-Brustel Y, Olichon-Berthe C, Grémeaux T, Tanti JF, Rochet N, Van Obberghen E. Glucose transporter in insulin sensitive tissues of lean and obese mice. Effect of the thermogenic agent BRL 26830A. Endocrinology. 1990 Dec;127(6):2687–2695. [PubMed] [Google Scholar]
- Li G, Stahl PD. Post-translational processing and membrane association of the two early endosome-associated rab GTP-binding proteins (rab4 and rab5). Arch Biochem Biophys. 1993 Aug 1;304(2):471–478. [PubMed] [Google Scholar]
- Li G, Stahl PD. Structure-function relationship of the small GTPase rab5. J Biol Chem. 1993 Nov 15;268(32):24475–24480. [PubMed] [Google Scholar]
- Livingstone C, James DE, Rice JE, Hanpeter D, Gould GW. Compartment ablation analysis of the insulin-responsive glucose transporter (GLUT4) in 3T3-L1 adipocytes. Biochem J. 1996 Apr 15;315(Pt 2):487–495. [PMC free article] [PubMed] [Google Scholar]
- Novick P, Brennwald P. Friends and family: the role of the Rab GTPases in vesicular traffic. Cell. 1993 Nov 19;75(4):597–601. [PubMed] [Google Scholar]
- Okada T, Masuda T, Shinkai M, Kariya K, Kataoka T. Post-translational modification of H-Ras is required for activation of, but not for association with, B-Raf. J Biol Chem. 1996 Mar 1;271(9):4671–4678. [PubMed] [Google Scholar]
- Quon MJ, Butte AJ, Zarnowski MJ, Sesti G, Cushman SW, Taylor SI. Insulin receptor substrate 1 mediates the stimulatory effect of insulin on GLUT4 translocation in transfected rat adipose cells. J Biol Chem. 1994 Nov 11;269(45):27920–27924. [PubMed] [Google Scholar]
- Quon MJ, Chen H, Ing BL, Liu ML, Zarnowski MJ, Yonezawa K, Kasuga M, Cushman SW, Taylor SI. Roles of 1-phosphatidylinositol 3-kinase and ras in regulating translocation of GLUT4 in transfected rat adipose cells. Mol Cell Biol. 1995 Oct;15(10):5403–5411. [PMC free article] [PubMed] [Google Scholar]
- Quon MJ, Guerre-Millo M, Zarnowski MJ, Butte AJ, Em M, Cushman SW, Taylor SI. Tyrosine kinase-deficient mutant human insulin receptors (Met1153-->Ile) overexpressed in transfected rat adipose cells fail to mediate translocation of epitope-tagged GLUT4. Proc Natl Acad Sci U S A. 1994 Jun 7;91(12):5587–5591. [PMC free article] [PubMed] [Google Scholar]
- Quon MJ, Zarnowski MJ, Guerre-Millo M, de la Luz Sierra M, Taylor SI, Cushman SW. Transfection of DNA into isolated rat adipose cells by electroporation: evaluation of promoter activity in transfected adipose cells which are highly responsive to insulin after one day in culture. Biochem Biophys Res Commun. 1993 Jul 15;194(1):338–346. [PubMed] [Google Scholar]
- Ricort JM, Tanti JF, Cormont M, Van Obberghen E, Le Marchand-Brustel Y. Parallel changes in Glut 4 and Rab4 movements in two insulin-resistant states. FEBS Lett. 1994 Jun 20;347(1):42–44. [PubMed] [Google Scholar]
- Robinson LJ, Pang S, Harris DS, Heuser J, James DE. Translocation of the glucose transporter (GLUT4) to the cell surface in permeabilized 3T3-L1 adipocytes: effects of ATP insulin, and GTP gamma S and localization of GLUT4 to clathrin lattices. J Cell Biol. 1992 Jun;117(6):1181–1196. [PMC free article] [PubMed] [Google Scholar]
- Rutter GA, Denton RM. Effects of insulin and guanosine 5'-[gamma-thio]triphosphate on fatty acid synthesis and lipolysis within electropermeabilized fat-cells. Biochem J. 1992 Jan 15;281(Pt 2):431–435. [PMC free article] [PubMed] [Google Scholar]
- Sherman LA, Hirshman MF, Cormont M, Le Marchand-Brustel Y, Goodyear LJ. Differential effects of insulin and exercise on Rab4 distribution in rat skeletal muscle. Endocrinology. 1996 Jan;137(1):266–273. [PubMed] [Google Scholar]
- Shibata H, Omata W, Suzuki Y, Tanaka S, Kojima I. A synthetic peptide corresponding to the Rab4 hypervariable carboxyl-terminal domain inhibits insulin action on glucose transport in rat adipocytes. J Biol Chem. 1996 Apr 19;271(16):9704–9709. [PubMed] [Google Scholar]
- Shibata H, Suzuki Y, Omata W, Tanaka S, Kojima I. Dissection of GLUT4 recycling pathway into exocytosis and endocytosis in rat adipocytes. Evidence that GTP-binding proteins are involved in both processes. J Biol Chem. 1995 May 12;270(19):11489–11495. [PubMed] [Google Scholar]
- Shirataki H, Kaibuchi K, Sakoda T, Kishida S, Yamaguchi T, Wada K, Miyazaki M, Takai Y. Rabphilin-3A, a putative target protein for smg p25A/rab3A p25 small GTP-binding protein related to synaptotagmin. Mol Cell Biol. 1993 Apr;13(4):2061–2068. [PMC free article] [PubMed] [Google Scholar]
- Simpson IA, Cushman SW. Hormonal regulation of mammalian glucose transport. Annu Rev Biochem. 1986;55:1059–1089. [PubMed] [Google Scholar]
- Slot JW, Geuze HJ, Gigengack S, James DE, Lienhard GE. Translocation of the glucose transporter GLUT4 in cardiac myocytes of the rat. Proc Natl Acad Sci U S A. 1991 Sep 1;88(17):7815–7819. [PMC free article] [PubMed] [Google Scholar]
- Slot JW, Geuze HJ, Gigengack S, Lienhard GE, James DE. Immuno-localization of the insulin regulatable glucose transporter in brown adipose tissue of the rat. J Cell Biol. 1991 Apr;113(1):123–135. [PMC free article] [PubMed] [Google Scholar]
- Stahl B, Chou JH, Li C, Südhof TC, Jahn R. Rab3 reversibly recruits rabphilin to synaptic vesicles by a mechanism analogous to raf recruitment by ras. EMBO J. 1996 Apr 15;15(8):1799–1809. [PMC free article] [PubMed] [Google Scholar]
- Stephens JM, Pilch PF. The metabolic regulation and vesicular transport of GLUT4, the major insulin-responsive glucose transporter. Endocr Rev. 1995 Aug;16(4):529–546. [PubMed] [Google Scholar]
- Tanti JF, Grémeaux T, Grillo S, Calleja V, Klippel A, Williams LT, Van Obberghen E, Le Marchand-Brustel Y. Overexpression of a constitutively active form of phosphatidylinositol 3-kinase is sufficient to promote Glut 4 translocation in adipocytes. J Biol Chem. 1996 Oct 11;271(41):25227–25232. [PubMed] [Google Scholar]
- Tozzo E, Kahn BB, Pilch PF, Kandror KV. Glut4 is targeted to specific vesicles in adipocytes of transgenic mice overexpressing Glut4 selectively in adipose tissue. J Biol Chem. 1996 May 3;271(18):10490–10494. [PubMed] [Google Scholar]
- Tozzo E, Shepherd PR, Gnudi L, Kahn BB. Transgenic GLUT-4 overexpression in fat enhances glucose metabolism: preferential effect on fatty acid synthesis. Am J Physiol. 1995 May;268(5 Pt 1):E956–E964. [PubMed] [Google Scholar]
- Tsao TS, Burcelin R, Katz EB, Huang L, Charron MJ. Enhanced insulin action due to targeted GLUT4 overexpression exclusively in muscle. Diabetes. 1996 Jan;45(1):28–36. [PubMed] [Google Scholar]
- Uphues I, Kolter T, Goud B, Eckel J. Insulin-induced translocation of the glucose transporter GLUT4 in cardiac muscle: studies on the role of small-molecular-mass GTP-binding proteins. Biochem J. 1994 Jul 1;301(Pt 1):177–182. [PMC free article] [PubMed] [Google Scholar]
- van der Sluijs P, Hull M, Webster P, Mâle P, Goud B, Mellman I. The small GTP-binding protein rab4 controls an early sorting event on the endocytic pathway. Cell. 1992 Sep 4;70(5):729–740. [PubMed] [Google Scholar]
- Van Der Sluijs P, Hull M, Zahraoui A, Tavitian A, Goud B, Mellman I. The small GTP-binding protein rab4 is associated with early endosomes. Proc Natl Acad Sci U S A. 1991 Jul 15;88(14):6313–6317. [PMC free article] [PubMed] [Google Scholar]
- Volchuk A, Sargeant R, Sumitani S, Liu Z, He L, Klip A. Cellubrevin is a resident protein of insulin-sensitive GLUT4 glucose transporter vesicles in 3T3-L1 adipocytes. J Biol Chem. 1995 Apr 7;270(14):8233–8240. [PubMed] [Google Scholar]
- Zahraoui A, Touchot N, Chardin P, Tavitian A. The human Rab genes encode a family of GTP-binding proteins related to yeast YPT1 and SEC4 products involved in secretion. J Biol Chem. 1989 Jul 25;264(21):12394–12401. [PubMed] [Google Scholar]
- Zerial M, Stenmark H. Rab GTPases in vesicular transport. Curr Opin Cell Biol. 1993 Aug;5(4):613–620. [PubMed] [Google Scholar]
- Zorzano A, Wilkinson W, Kotliar N, Thoidis G, Wadzinkski BE, Ruoho AE, Pilch PF. Insulin-regulated glucose uptake in rat adipocytes is mediated by two transporter isoforms present in at least two vesicle populations. J Biol Chem. 1989 Jul 25;264(21):12358–12363. [PubMed] [Google Scholar]