Rip11 is a Rab11- and AS160-RabGAP-binding protein required for insulin-stimulated glucose uptake in adipocytes.
The translocation of GLUT4 to the plasma membrane underlies the ability of insulin to stimulate glucose uptake, an event that involves the activation of protein kinase B, several members of the Rab family of GTP-binding proteins and the phosphorylation of the Rab GTPase-activating protein AS160. Here, we explored the regulation by insulin of the class I Rab11-interacting proteins Rip11, RCP and FIP2. We show that Rip11, but not RCP or FIP2, translocates to the plasma membrane of 3T3-L1 adipocytes in response to insulin. This unique response of Rip11 prompted us to explore the role of this protein in more detail. We found that Rip11 partially colocalises with GLUT4 in intracellular compartments. siRNA-mediated knockdown of Rip11 inhibits insulin-stimulated uptake of 2-deoxyglucose, and overexpression of Rip11 blocks insulin-stimulated insertion of translocated GLUT4 vesicles into the plasma membrane. We additionally show that Rip11 forms a complex with AS160 in a Rab11-independent manner and that insulin induces dissociation of AS160 from Rip11. We propose that Rip11 is an AS160- and Rab-binding protein that coordinates the protein kinase signalling and trafficking machinery required to stimulate glucose uptake in response to insulin.
Welsh
Gavin I
GI
Department of Biochemistry, School of Medical Sciences, University of Bristol, Bristol, BS8 ITD, UK.
Leney
Sophie E
SE
Lloyd-Lewis
Bethan
B
Wherlock
Matthew
M
Lindsay
Andrew J
AJ
McCaffrey
Mary W
MW
Tavaré
Jeremy M
JM
D041721:3T3-L1 Cells; D017667:Adipocytes / Q000166:cytology / Q000378:metabolism*; D000818:Animals; D016466:CHO Cells; D002352:Carrier Proteins; D002454:Cell Differentiation; D002462:Cell Membrane / Q000378:metabolism; D002999:Clone Cells; D006224:Cricetinae; D003412:Cricetulus; D003847:Deoxyglucose / Q000037:antagonists & inhibitors; D018274:Electroporation; D005347:Fibroblasts / Q000378:metabolism; D019084:Fluorescent Antibody Technique, Indirect; D005947:Glucose / Q000378:metabolism*; D051275:Glucose Transporter Type 4 / Q000235:genetics / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000378:metabolism; D007328:Insulin / Q000494:pharmacology*; D051379:Mice; D024101:Mitochondrial Proteins; D008954:Models, Biological; D011233:Precipitin Tests; D021381:Protein Transport / Q000187:drug effects; D034741:RNA, Small Interfering / Q000378:metabolism; D013997:Time Factors; D014162:Transfection; D020691:rab GTP-Binding Proteins / Q000235:genetics / Q000502:physiology*
D002352:Carrier Proteins; D051275:Glucose Transporter Type 4; D007328:Insulin; D024101:Mitochondrial Proteins; D034741:RNA, Small Interfering; C527553:Rip11 protein, mouse; D049452:Green Fluorescent Proteins; D003847:Deoxyglucose; D020691:rab GTP-Binding Proteins; D005947:Glucose