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PMID: 12832475 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Insulin-induced GLUT4 translocation involves protein kinase C-lambda-mediated functional coupling between Rab4 and the motor protein kinesin.

Molecular and cellular biology ·Vol. 23 ·No. 14 ·2003-07-00 ·Pages 4892-900

Imamura T, Huang J, Usui I, Satoh H, Bever J, Olefsky JM

Abstract

Insulin stimulates glucose transport by promoting translocation of GLUT4 proteins from the perinuclear compartment to the cell surface. It has been previously suggested that the microtubule-associated motor protein kinesin, which transports cargo toward the plus end of microtubules, plays a role in translocating GLUT4 vesicles to the cell surface. In this study, we investigated the role of Rab4, a small GTPase-binding protein, and the motor protein KIF3 (kinesin II in mice) in insulin-induced GLUT4 exocytosis in 3T3-L1 adipocytes. Photoaffinity labeling of Rab4 with [gamma-(32)P]GTP-azidoanilide showed that insulin stimulated Rab4 GTP loading and that this insulin effect was inhibited by pretreatment with the phosphatidylinositol 3-kinase (PI3-kinase) inhibitor LY294002 or expression of dominant-negative protein kinase C-lambda (PKC-lambda). Consistent with previous reports, expression of dominant-negative Rab4 (N121I) decreased insulin-induced GLUT4 translocation by 45%. Microinjection of an anti-KIF3 antibody into 3T3-L1 adipocytes decreased insulin-induced GLUT4 exocytosis by 65% but had no effect on endocytosis. Coimmunoprecipitation experiments showed that Rab4, but not Rab5, physically associated with KIF3, and this was confirmed by showing in vitro association using glutathione S-transferase-Rab4. A microtubule capture assay demonstrated that insulin stimulation increased the activity for the binding of KIF3 to microtubules and that this activation was inhibited by pretreatment with the PI3-kinase inhibitor LY294002 or expression of dominant-negative PKC-lambda. Taken together, these data indicate that (i) insulin signaling stimulates Rab4 activity, the association of Rab4 with kinesin, and the interaction of KIF3 with microtubules and (ii) this process is mediated by insulin-induced PI3-kinase-dependent PKC-lambda activation and participates in GLUT4 exocytosis in 3T3-L1 adipocytes.

MeSH Terms
3T3 Cells/drug effects,metabolism Adipocytes/drug effects,metabolism Animals Biological Assay/methods Cell Membrane/metabolism Chromones/pharmacology Enzyme Activation/drug effects Enzyme Inhibitors/pharmacology Exocytosis/physiology Glucose Transporter Type 4 Insulin/metabolism,pharmacology Isoenzymes Kinesins/genetics,metabolism Mice Microtubules/metabolism Monosaccharide Transport Proteins/genetics,metabolism Morpholines/pharmacology Muscle Proteins Phosphatidylinositol 3-Kinases/drug effects,metabolism Phosphoinositide-3 Kinase Inhibitors Protein Kinase C/genetics,metabolism Protein Transport/drug effects Signal Transduction Transport Vesicles/metabolism rab4 GTP-Binding Proteins/genetics,metabolism rab5 GTP-Binding Proteins/drug effects,metabolism
Chemicals
Chromones Enzyme Inhibitors Glucose Transporter Type 4 Insulin Isoenzymes Kif3a protein, mouse Monosaccharide Transport Proteins Morpholines Muscle Proteins Phosphoinositide-3 Kinase Inhibitors Slc2a4 protein, mouse 2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one Protein Kinase C protein kinase C lambda Kinesins rab4 GTP-Binding Proteins rab5 GTP-Binding Proteins
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Imamura Takeshi
Division of Endocrinology and Metabolism, Department of Medicine, University of California, San Diego, La Jolla, California 92093, USA.
Huang Jie
Usui Isao
Satoh Hiroaki
Bever Jennie
Olefsky Jerrold M
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.23.14.4892-4900.2003
Published
2003-07-00
Pages
4892-900
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC162221
Subset
IM
Grants
NIDDK NIH HHS · R01 DK033651 · United States
NIDDK NIH HHS · R37 DK033651 · United States
NIDDK NIH HHS · DK-33651 · United States
External Links
PubMed source DOI: 10.1128/MCB.23.14.4892-4900.2003 PMC Full Text View journal details
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