Integrin-linked kinase is an essential mediator for T-cadherin-dependent signaling via Akt and GSK3β in endothelial cells

Manjunath B. Joshi, Danila Ivanov, Maria Philippova, Paul Erne, Thérèse J. Resink

Research output: Contribution to journalArticle

45 Citations (Scopus)

Abstract

Glycosylphosphatidylinositol-anchored T-cadherin (T-cad) influences several parameters of angiogenesis including endothelial cell (EC) differentiation, migration, proliferation, and survival. This presupposes signal transduction networking via mediatory regulators and molecular adaptors since T-cad lacks transmembrane and cytosolic domains. Here, using pharmacological inhibition of PI3K, adenoviral-mediated T-cad-overexpression, siRNA-mediated T-cad-depletion, and agonistic antibody-mediated ligation, we demonstrate signaling by T-cad through PI3K-Akt-GSK3β pathways in EC. T-cad-overexpressing EC exhibited increased levels and nuclear accumulation of active β-catenin, which was transcriptionally active as shown by increased Lef/Tcf reporter activity and cyclin D1 levels. Cotransduction of EC with constitutively active GSK3β (S9A-GSK3β) abrogated the stimulatory effects of T-cad on active β-catenin accumulation, proliferation, and survival. Integrin-linked kinase (ILK), a membrane proximal upstream regulator of Akt and GSK3β, was considered a candidate signaling mediator for T-cad. T-cad was present in anti-ILK immunoprecipitates, and confocal microscopy revealed colocalization of T-cad and ILK within lamellipodia of migrating cells. ILK-siRNA abolished T-cad-dependent effects on Ser-473Akt/Ser-9GSK3β phosphorylation, active β-catenin accumulation, and survival. We conclude ILK is an essential mediator for T-cad signaling via Akt and GSK3β in EC. This is the first demonstration that ILK can regulate inward signaling by GPI-anchored proteins. Furthermore, ILK-GSK3β-dependent modulation of active β-catenin levels by GPI-anchored T-cad represents a novel mechanism for controlling cellular β-catenin activity.

Original languageEnglish
Pages (from-to)3083-3095
Number of pages13
JournalFASEB Journal
Volume21
Issue number12
DOIs
Publication statusPublished - 10-2007

Fingerprint

cadherins
Endothelial cells
integrins
endothelial cells
Catenins
phosphotransferases (kinases)
Endothelial Cells
phosphatidylinositol 3-kinase
small interfering RNA
Phosphatidylinositol 3-Kinases
Small Interfering RNA
pseudopodia
Glycosylphosphatidylinositols
Signal transduction
Phosphorylation
Pseudopodia
Confocal microscopy
Cyclin D1
cyclins
angiogenesis

All Science Journal Classification (ASJC) codes

  • Agricultural and Biological Sciences (miscellaneous)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Cell Biology
  • Medicine(all)

Cite this

Joshi, Manjunath B. ; Ivanov, Danila ; Philippova, Maria ; Erne, Paul ; Resink, Thérèse J. / Integrin-linked kinase is an essential mediator for T-cadherin-dependent signaling via Akt and GSK3β in endothelial cells. In: FASEB Journal. 2007 ; Vol. 21, No. 12. pp. 3083-3095.
@article{0692f666815746ebb3723dec00dd3eec,
title = "Integrin-linked kinase is an essential mediator for T-cadherin-dependent signaling via Akt and GSK3β in endothelial cells",
abstract = "Glycosylphosphatidylinositol-anchored T-cadherin (T-cad) influences several parameters of angiogenesis including endothelial cell (EC) differentiation, migration, proliferation, and survival. This presupposes signal transduction networking via mediatory regulators and molecular adaptors since T-cad lacks transmembrane and cytosolic domains. Here, using pharmacological inhibition of PI3K, adenoviral-mediated T-cad-overexpression, siRNA-mediated T-cad-depletion, and agonistic antibody-mediated ligation, we demonstrate signaling by T-cad through PI3K-Akt-GSK3β pathways in EC. T-cad-overexpressing EC exhibited increased levels and nuclear accumulation of active β-catenin, which was transcriptionally active as shown by increased Lef/Tcf reporter activity and cyclin D1 levels. Cotransduction of EC with constitutively active GSK3β (S9A-GSK3β) abrogated the stimulatory effects of T-cad on active β-catenin accumulation, proliferation, and survival. Integrin-linked kinase (ILK), a membrane proximal upstream regulator of Akt and GSK3β, was considered a candidate signaling mediator for T-cad. T-cad was present in anti-ILK immunoprecipitates, and confocal microscopy revealed colocalization of T-cad and ILK within lamellipodia of migrating cells. ILK-siRNA abolished T-cad-dependent effects on Ser-473Akt/Ser-9GSK3β phosphorylation, active β-catenin accumulation, and survival. We conclude ILK is an essential mediator for T-cad signaling via Akt and GSK3β in EC. This is the first demonstration that ILK can regulate inward signaling by GPI-anchored proteins. Furthermore, ILK-GSK3β-dependent modulation of active β-catenin levels by GPI-anchored T-cad represents a novel mechanism for controlling cellular β-catenin activity.",
author = "Joshi, {Manjunath B.} and Danila Ivanov and Maria Philippova and Paul Erne and Resink, {Th{\'e}r{\`e}se J.}",
year = "2007",
month = "10",
doi = "10.1096/fj.06-7723com",
language = "English",
volume = "21",
pages = "3083--3095",
journal = "FASEB Journal",
issn = "0892-6638",
publisher = "FASEB",
number = "12",

}

Integrin-linked kinase is an essential mediator for T-cadherin-dependent signaling via Akt and GSK3β in endothelial cells. / Joshi, Manjunath B.; Ivanov, Danila; Philippova, Maria; Erne, Paul; Resink, Thérèse J.

In: FASEB Journal, Vol. 21, No. 12, 10.2007, p. 3083-3095.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Integrin-linked kinase is an essential mediator for T-cadherin-dependent signaling via Akt and GSK3β in endothelial cells

AU - Joshi, Manjunath B.

AU - Ivanov, Danila

AU - Philippova, Maria

AU - Erne, Paul

AU - Resink, Thérèse J.

PY - 2007/10

Y1 - 2007/10

N2 - Glycosylphosphatidylinositol-anchored T-cadherin (T-cad) influences several parameters of angiogenesis including endothelial cell (EC) differentiation, migration, proliferation, and survival. This presupposes signal transduction networking via mediatory regulators and molecular adaptors since T-cad lacks transmembrane and cytosolic domains. Here, using pharmacological inhibition of PI3K, adenoviral-mediated T-cad-overexpression, siRNA-mediated T-cad-depletion, and agonistic antibody-mediated ligation, we demonstrate signaling by T-cad through PI3K-Akt-GSK3β pathways in EC. T-cad-overexpressing EC exhibited increased levels and nuclear accumulation of active β-catenin, which was transcriptionally active as shown by increased Lef/Tcf reporter activity and cyclin D1 levels. Cotransduction of EC with constitutively active GSK3β (S9A-GSK3β) abrogated the stimulatory effects of T-cad on active β-catenin accumulation, proliferation, and survival. Integrin-linked kinase (ILK), a membrane proximal upstream regulator of Akt and GSK3β, was considered a candidate signaling mediator for T-cad. T-cad was present in anti-ILK immunoprecipitates, and confocal microscopy revealed colocalization of T-cad and ILK within lamellipodia of migrating cells. ILK-siRNA abolished T-cad-dependent effects on Ser-473Akt/Ser-9GSK3β phosphorylation, active β-catenin accumulation, and survival. We conclude ILK is an essential mediator for T-cad signaling via Akt and GSK3β in EC. This is the first demonstration that ILK can regulate inward signaling by GPI-anchored proteins. Furthermore, ILK-GSK3β-dependent modulation of active β-catenin levels by GPI-anchored T-cad represents a novel mechanism for controlling cellular β-catenin activity.

AB - Glycosylphosphatidylinositol-anchored T-cadherin (T-cad) influences several parameters of angiogenesis including endothelial cell (EC) differentiation, migration, proliferation, and survival. This presupposes signal transduction networking via mediatory regulators and molecular adaptors since T-cad lacks transmembrane and cytosolic domains. Here, using pharmacological inhibition of PI3K, adenoviral-mediated T-cad-overexpression, siRNA-mediated T-cad-depletion, and agonistic antibody-mediated ligation, we demonstrate signaling by T-cad through PI3K-Akt-GSK3β pathways in EC. T-cad-overexpressing EC exhibited increased levels and nuclear accumulation of active β-catenin, which was transcriptionally active as shown by increased Lef/Tcf reporter activity and cyclin D1 levels. Cotransduction of EC with constitutively active GSK3β (S9A-GSK3β) abrogated the stimulatory effects of T-cad on active β-catenin accumulation, proliferation, and survival. Integrin-linked kinase (ILK), a membrane proximal upstream regulator of Akt and GSK3β, was considered a candidate signaling mediator for T-cad. T-cad was present in anti-ILK immunoprecipitates, and confocal microscopy revealed colocalization of T-cad and ILK within lamellipodia of migrating cells. ILK-siRNA abolished T-cad-dependent effects on Ser-473Akt/Ser-9GSK3β phosphorylation, active β-catenin accumulation, and survival. We conclude ILK is an essential mediator for T-cad signaling via Akt and GSK3β in EC. This is the first demonstration that ILK can regulate inward signaling by GPI-anchored proteins. Furthermore, ILK-GSK3β-dependent modulation of active β-catenin levels by GPI-anchored T-cad represents a novel mechanism for controlling cellular β-catenin activity.

UR - http://www.scopus.com/inward/record.url?scp=35248896777&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=35248896777&partnerID=8YFLogxK

U2 - 10.1096/fj.06-7723com

DO - 10.1096/fj.06-7723com

M3 - Article

VL - 21

SP - 3083

EP - 3095

JO - FASEB Journal

JF - FASEB Journal

SN - 0892-6638

IS - 12

ER -