The integrin cytoplasmic domain-associated protein ICAP-1 binds and regulates Rho family GTPases during cell spreading

Simona Degani, Fiorella Balzac, Mara Brancaccio, Simona Guazzone, Saverio Francesco Retta, Lorenzo Silengo, Alessandra Eva, Guido Tarone

Research output: Contribution to journalArticle

47 Citations (Scopus)

Abstract

Using two-hybrid screening, we isolated the integrin cytoplasmic domain-associated protein (ICAP-1), an interactor for the COOH terminal region of the β1A integrin cytoplasmic domain. To investigate the role of ICAP-1 in integrin-mediated adhesive function, we expressed the full-length molecule in NIH3T3 cells. ICAP-1 expression strongly prevents NIH3T3 cell spreading on extracellular matrix. This inhibition is transient and can be counteracted by coexpression of a constitutively activated mutant of Cdc42, suggesting that ICAP-1 acts upstream of this GTPase. In addition, we found that ICAP-1 binds both to Cdc42 and Rac1 in vitro, and its expression markedly inhibits activation of these GTPases during integrin-mediated cell adhesion to fibronectin as detected by PAK binding assay. In the attempt to define the molecular mechanism of this inhibition, we show that ICAP-1 reduces both the intrinsic and the exchange factor-induced dissociation of GDP from Cdc42; moreover, purified ICAP-1 displaces this GTPase from cellular membranes. Together, these data show for the first time that ICAP-1 regulates Rho family GTPases during integrin-mediated cell matrix adhesion, acting as guanine dissociation inhibitor.

Original languageEnglish
Pages (from-to)377-387
Number of pages11
JournalJournal of Cell Biology
Volume156
Issue number2
DOIs
Publication statusPublished - Jan 21 2002

Fingerprint

rho GTP-Binding Proteins
Integrins
GTP Phosphohydrolases
Cell-Matrix Junctions
Intrinsic Factor
Guanine
Fibronectins
Cell Adhesion
Adhesives
Extracellular Matrix
Protein Domains
Membranes

Keywords

  • Cdc42
  • Cytoskeleton
  • GDI
  • ICAP-1
  • Integrins
  • Rac

ASJC Scopus subject areas

  • Cell Biology

Cite this

Degani, S., Balzac, F., Brancaccio, M., Guazzone, S., Retta, S. F., Silengo, L., ... Tarone, G. (2002). The integrin cytoplasmic domain-associated protein ICAP-1 binds and regulates Rho family GTPases during cell spreading. Journal of Cell Biology, 156(2), 377-387. https://doi.org/10.1083/jcb.200108030

The integrin cytoplasmic domain-associated protein ICAP-1 binds and regulates Rho family GTPases during cell spreading. / Degani, Simona; Balzac, Fiorella; Brancaccio, Mara; Guazzone, Simona; Retta, Saverio Francesco; Silengo, Lorenzo; Eva, Alessandra; Tarone, Guido.

In: Journal of Cell Biology, Vol. 156, No. 2, 21.01.2002, p. 377-387.

Research output: Contribution to journalArticle

Degani, S, Balzac, F, Brancaccio, M, Guazzone, S, Retta, SF, Silengo, L, Eva, A & Tarone, G 2002, 'The integrin cytoplasmic domain-associated protein ICAP-1 binds and regulates Rho family GTPases during cell spreading', Journal of Cell Biology, vol. 156, no. 2, pp. 377-387. https://doi.org/10.1083/jcb.200108030
Degani, Simona ; Balzac, Fiorella ; Brancaccio, Mara ; Guazzone, Simona ; Retta, Saverio Francesco ; Silengo, Lorenzo ; Eva, Alessandra ; Tarone, Guido. / The integrin cytoplasmic domain-associated protein ICAP-1 binds and regulates Rho family GTPases during cell spreading. In: Journal of Cell Biology. 2002 ; Vol. 156, No. 2. pp. 377-387.
@article{5462c7635bae42cca01558699f7f8803,
title = "The integrin cytoplasmic domain-associated protein ICAP-1 binds and regulates Rho family GTPases during cell spreading",
abstract = "Using two-hybrid screening, we isolated the integrin cytoplasmic domain-associated protein (ICAP-1), an interactor for the COOH terminal region of the β1A integrin cytoplasmic domain. To investigate the role of ICAP-1 in integrin-mediated adhesive function, we expressed the full-length molecule in NIH3T3 cells. ICAP-1 expression strongly prevents NIH3T3 cell spreading on extracellular matrix. This inhibition is transient and can be counteracted by coexpression of a constitutively activated mutant of Cdc42, suggesting that ICAP-1 acts upstream of this GTPase. In addition, we found that ICAP-1 binds both to Cdc42 and Rac1 in vitro, and its expression markedly inhibits activation of these GTPases during integrin-mediated cell adhesion to fibronectin as detected by PAK binding assay. In the attempt to define the molecular mechanism of this inhibition, we show that ICAP-1 reduces both the intrinsic and the exchange factor-induced dissociation of GDP from Cdc42; moreover, purified ICAP-1 displaces this GTPase from cellular membranes. Together, these data show for the first time that ICAP-1 regulates Rho family GTPases during integrin-mediated cell matrix adhesion, acting as guanine dissociation inhibitor.",
keywords = "Cdc42, Cytoskeleton, GDI, ICAP-1, Integrins, Rac",
author = "Simona Degani and Fiorella Balzac and Mara Brancaccio and Simona Guazzone and Retta, {Saverio Francesco} and Lorenzo Silengo and Alessandra Eva and Guido Tarone",
year = "2002",
month = "1",
day = "21",
doi = "10.1083/jcb.200108030",
language = "English",
volume = "156",
pages = "377--387",
journal = "Journal of Cell Biology",
issn = "0021-9525",
publisher = "Rockefeller University Press",
number = "2",

}

TY - JOUR

T1 - The integrin cytoplasmic domain-associated protein ICAP-1 binds and regulates Rho family GTPases during cell spreading

AU - Degani, Simona

AU - Balzac, Fiorella

AU - Brancaccio, Mara

AU - Guazzone, Simona

AU - Retta, Saverio Francesco

AU - Silengo, Lorenzo

AU - Eva, Alessandra

AU - Tarone, Guido

PY - 2002/1/21

Y1 - 2002/1/21

N2 - Using two-hybrid screening, we isolated the integrin cytoplasmic domain-associated protein (ICAP-1), an interactor for the COOH terminal region of the β1A integrin cytoplasmic domain. To investigate the role of ICAP-1 in integrin-mediated adhesive function, we expressed the full-length molecule in NIH3T3 cells. ICAP-1 expression strongly prevents NIH3T3 cell spreading on extracellular matrix. This inhibition is transient and can be counteracted by coexpression of a constitutively activated mutant of Cdc42, suggesting that ICAP-1 acts upstream of this GTPase. In addition, we found that ICAP-1 binds both to Cdc42 and Rac1 in vitro, and its expression markedly inhibits activation of these GTPases during integrin-mediated cell adhesion to fibronectin as detected by PAK binding assay. In the attempt to define the molecular mechanism of this inhibition, we show that ICAP-1 reduces both the intrinsic and the exchange factor-induced dissociation of GDP from Cdc42; moreover, purified ICAP-1 displaces this GTPase from cellular membranes. Together, these data show for the first time that ICAP-1 regulates Rho family GTPases during integrin-mediated cell matrix adhesion, acting as guanine dissociation inhibitor.

AB - Using two-hybrid screening, we isolated the integrin cytoplasmic domain-associated protein (ICAP-1), an interactor for the COOH terminal region of the β1A integrin cytoplasmic domain. To investigate the role of ICAP-1 in integrin-mediated adhesive function, we expressed the full-length molecule in NIH3T3 cells. ICAP-1 expression strongly prevents NIH3T3 cell spreading on extracellular matrix. This inhibition is transient and can be counteracted by coexpression of a constitutively activated mutant of Cdc42, suggesting that ICAP-1 acts upstream of this GTPase. In addition, we found that ICAP-1 binds both to Cdc42 and Rac1 in vitro, and its expression markedly inhibits activation of these GTPases during integrin-mediated cell adhesion to fibronectin as detected by PAK binding assay. In the attempt to define the molecular mechanism of this inhibition, we show that ICAP-1 reduces both the intrinsic and the exchange factor-induced dissociation of GDP from Cdc42; moreover, purified ICAP-1 displaces this GTPase from cellular membranes. Together, these data show for the first time that ICAP-1 regulates Rho family GTPases during integrin-mediated cell matrix adhesion, acting as guanine dissociation inhibitor.

KW - Cdc42

KW - Cytoskeleton

KW - GDI

KW - ICAP-1

KW - Integrins

KW - Rac

UR - http://www.scopus.com/inward/record.url?scp=0037148525&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0037148525&partnerID=8YFLogxK

U2 - 10.1083/jcb.200108030

DO - 10.1083/jcb.200108030

M3 - Article

C2 - 11807099

AN - SCOPUS:0037148525

VL - 156

SP - 377

EP - 387

JO - Journal of Cell Biology

JF - Journal of Cell Biology

SN - 0021-9525

IS - 2

ER -