Expression of a dominant interfering dynamin mutant in 3T3L1 adipocytes inhibits GLUT4 endocytosis without affecting insulin signaling

Aimee W. Kao, Brian P. Ceresa, Scott R. Santeler, Jeffrey E. Pessin

Research output: Contribution to journalArticle

104 Citations (Scopus)

Abstract

To examine the role of clathrin-coated vesicle endocytosis in insulin receptor signaling and GLUT4 trafficking, we used recombinant adenovirus to express a dominant interfering mutant of dynamin (K44A/dynamin) in 3T3L1 adipocytes. Functional expression of K44A/dynamin, as measured by inhibition of transferrin receptor internalization, did not affect insulin-stimulated insulin receptor autophosphorylation, Shc tyrosine phosphorylation, or mitogen-activated protein kinase activation. Although the tyrosine phosphorylation of insulin receptor substrate-1 was slightly reduced, correlating with a 25% decrease in insulin receptor substrate-1-associated phosphatidylinositol 3-kinase activity, insulin-stimulated Akt kinase activation was unaffected. In contrast, expression of K44A/dynamin resulted in the cell-surface accumulation of GLUT4 under basal conditions and an inhibition of GLUT4 endocytosis without affecting insulin-stimulated GLUT4 exocytosis. These data demonstrate that disruption of clathrin-mediated endocytosis does not significantly perturb insulin receptor signal transduction pathways. Furthermore, K44A/dynamin expression causes an accumulation of GLUT4 at the cell surface, suggesting that GLUT4 vesicles exist in at least two distinct intracellular compartments, one that undergoes continuous recycling and a second that is responsive to insulin.

Original languageEnglish (US)
Pages (from-to)25450-25457
Number of pages8
JournalJournal of Biological Chemistry
Volume273
Issue number39
DOIs
StatePublished - Sep 25 1998
Externally publishedYes

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Dynamins
Endocytosis
Adipocytes
Insulin
Insulin Receptor
Insulin Receptor Substrate Proteins
Clathrin
Phosphorylation
Tyrosine
Chemical activation
Clathrin-Coated Vesicles
Phosphatidylinositol 3-Kinase
Signal transduction
Transferrin Receptors
Exocytosis
Recycling
Mitogen-Activated Protein Kinases
Adenoviridae
Signal Transduction
Phosphotransferases

ASJC Scopus subject areas

  • Biochemistry

Cite this

Expression of a dominant interfering dynamin mutant in 3T3L1 adipocytes inhibits GLUT4 endocytosis without affecting insulin signaling. / Kao, Aimee W.; Ceresa, Brian P.; Santeler, Scott R.; Pessin, Jeffrey E.

In: Journal of Biological Chemistry, Vol. 273, No. 39, 25.09.1998, p. 25450-25457.

Research output: Contribution to journalArticle

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