Inhibition of clathrin-mediated endocytosis selectively attenuates specific insulin receptor signal transduction pathways

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

Research output: Contribution to journalArticle

182 Citations (Scopus)

Abstract

To examine the role of clathrin-dependent insulin receptor internalization in insulin, stimulated signal transduction events, we expressed a dominant-interfering mutant of dynamin (K44A/dynamin) by using a recombinant adenovirus in the H4IIE hepatoma and 3T3L1 adipocyte-cell lines. Expression of K44A/dynamin inhibited endocytosis of the insulin receptor as determined by both cell surface radioligand binding and trypsin protection analysis. The inhibition of the insulin receptor endocytosis had no effect on either the extent of insulin receptor autophosphorylation or insulin receptor substrate 2 (IRS1) tyrosine phosphorylation contrast, expression of K44A/dynamin partially inhibited insulin-stimulated Shc tyrosine phosphorylation and activation of the mitogen-activated protein kinases ERK1 and -2. Although there was an approximately 50% decrease in the insulin- stimulated activation of the phosphatidylinositol 3-kinase associated with IRS1, insulin-stimulated Akt kinase phosphorylation and activation were unaffected. The expression of K44A/dynamin increased the basal rate of amino acid transport, which was additive with the effect of insulin but had no effect on the basal or insulin-stimulated DNA synthesis. In 3T3L1 adipocytes, expression of K44A/dynamin increased the basal rate of glucose uptake, glycogen synthesis, and lipogenesis without any significant effect on insulin stimulation. Together, these data demonstrate that the acute actions of insulin are largely independent of insulin receptor endocytosis and are initiated by activation of the plasma membrane-localized insulin receptor.

Original languageEnglish (US)
Pages (from-to)3862-3870
Number of pages9
JournalMolecular and Cellular Biology
Volume18
Issue number7
StatePublished - Jul 1998
Externally publishedYes

Fingerprint

Clathrin
Insulin Receptor
Endocytosis
Dynamins
Signal Transduction
Insulin
Phosphorylation
Adipocytes
Phosphatidylinositol 3-Kinase
Insulin Receptor Substrate Proteins
Lipogenesis
Mitogen-Activated Protein Kinase 1
Glycogen
Adenoviridae
Trypsin
Tyrosine
Hepatocellular Carcinoma
Phosphotransferases
Cell Membrane
Amino Acids

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cell Biology

Cite this

Inhibition of clathrin-mediated endocytosis selectively attenuates specific insulin receptor signal transduction pathways. / Ceresa, Brian P.; Kao, Aimee W.; Santeler, Scott R.; Pessin, Jeffrey E.

In: Molecular and Cellular Biology, Vol. 18, No. 7, 07.1998, p. 3862-3870.

Research output: Contribution to journalArticle

@article{c55f4ad09a244aa48a1a9735d0d07e78,
title = "Inhibition of clathrin-mediated endocytosis selectively attenuates specific insulin receptor signal transduction pathways",
abstract = "To examine the role of clathrin-dependent insulin receptor internalization in insulin, stimulated signal transduction events, we expressed a dominant-interfering mutant of dynamin (K44A/dynamin) by using a recombinant adenovirus in the H4IIE hepatoma and 3T3L1 adipocyte-cell lines. Expression of K44A/dynamin inhibited endocytosis of the insulin receptor as determined by both cell surface radioligand binding and trypsin protection analysis. The inhibition of the insulin receptor endocytosis had no effect on either the extent of insulin receptor autophosphorylation or insulin receptor substrate 2 (IRS1) tyrosine phosphorylation contrast, expression of K44A/dynamin partially inhibited insulin-stimulated Shc tyrosine phosphorylation and activation of the mitogen-activated protein kinases ERK1 and -2. Although there was an approximately 50{\%} decrease in the insulin- stimulated activation of the phosphatidylinositol 3-kinase associated with IRS1, insulin-stimulated Akt kinase phosphorylation and activation were unaffected. The expression of K44A/dynamin increased the basal rate of amino acid transport, which was additive with the effect of insulin but had no effect on the basal or insulin-stimulated DNA synthesis. In 3T3L1 adipocytes, expression of K44A/dynamin increased the basal rate of glucose uptake, glycogen synthesis, and lipogenesis without any significant effect on insulin stimulation. Together, these data demonstrate that the acute actions of insulin are largely independent of insulin receptor endocytosis and are initiated by activation of the plasma membrane-localized insulin receptor.",
author = "Ceresa, {Brian P.} and Kao, {Aimee W.} and Santeler, {Scott R.} and Pessin, {Jeffrey E.}",
year = "1998",
month = "7",
language = "English (US)",
volume = "18",
pages = "3862--3870",
journal = "Molecular and Cellular Biology",
issn = "0270-7306",
publisher = "American Society for Microbiology",
number = "7",

}

TY - JOUR

T1 - Inhibition of clathrin-mediated endocytosis selectively attenuates specific insulin receptor signal transduction pathways

AU - Ceresa, Brian P.

AU - Kao, Aimee W.

AU - Santeler, Scott R.

AU - Pessin, Jeffrey E.

PY - 1998/7

Y1 - 1998/7

N2 - To examine the role of clathrin-dependent insulin receptor internalization in insulin, stimulated signal transduction events, we expressed a dominant-interfering mutant of dynamin (K44A/dynamin) by using a recombinant adenovirus in the H4IIE hepatoma and 3T3L1 adipocyte-cell lines. Expression of K44A/dynamin inhibited endocytosis of the insulin receptor as determined by both cell surface radioligand binding and trypsin protection analysis. The inhibition of the insulin receptor endocytosis had no effect on either the extent of insulin receptor autophosphorylation or insulin receptor substrate 2 (IRS1) tyrosine phosphorylation contrast, expression of K44A/dynamin partially inhibited insulin-stimulated Shc tyrosine phosphorylation and activation of the mitogen-activated protein kinases ERK1 and -2. Although there was an approximately 50% decrease in the insulin- stimulated activation of the phosphatidylinositol 3-kinase associated with IRS1, insulin-stimulated Akt kinase phosphorylation and activation were unaffected. The expression of K44A/dynamin increased the basal rate of amino acid transport, which was additive with the effect of insulin but had no effect on the basal or insulin-stimulated DNA synthesis. In 3T3L1 adipocytes, expression of K44A/dynamin increased the basal rate of glucose uptake, glycogen synthesis, and lipogenesis without any significant effect on insulin stimulation. Together, these data demonstrate that the acute actions of insulin are largely independent of insulin receptor endocytosis and are initiated by activation of the plasma membrane-localized insulin receptor.

AB - To examine the role of clathrin-dependent insulin receptor internalization in insulin, stimulated signal transduction events, we expressed a dominant-interfering mutant of dynamin (K44A/dynamin) by using a recombinant adenovirus in the H4IIE hepatoma and 3T3L1 adipocyte-cell lines. Expression of K44A/dynamin inhibited endocytosis of the insulin receptor as determined by both cell surface radioligand binding and trypsin protection analysis. The inhibition of the insulin receptor endocytosis had no effect on either the extent of insulin receptor autophosphorylation or insulin receptor substrate 2 (IRS1) tyrosine phosphorylation contrast, expression of K44A/dynamin partially inhibited insulin-stimulated Shc tyrosine phosphorylation and activation of the mitogen-activated protein kinases ERK1 and -2. Although there was an approximately 50% decrease in the insulin- stimulated activation of the phosphatidylinositol 3-kinase associated with IRS1, insulin-stimulated Akt kinase phosphorylation and activation were unaffected. The expression of K44A/dynamin increased the basal rate of amino acid transport, which was additive with the effect of insulin but had no effect on the basal or insulin-stimulated DNA synthesis. In 3T3L1 adipocytes, expression of K44A/dynamin increased the basal rate of glucose uptake, glycogen synthesis, and lipogenesis without any significant effect on insulin stimulation. Together, these data demonstrate that the acute actions of insulin are largely independent of insulin receptor endocytosis and are initiated by activation of the plasma membrane-localized insulin receptor.

UR - http://www.scopus.com/inward/record.url?scp=0031806620&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0031806620&partnerID=8YFLogxK

M3 - Article

VL - 18

SP - 3862

EP - 3870

JO - Molecular and Cellular Biology

JF - Molecular and Cellular Biology

SN - 0270-7306

IS - 7

ER -