Steel factor enhances integrin-mediated tyrosine phosphorylation of focal adhesion kinase (pp125(FAK)) and paxillin

Hiroyuki Takahira, Akihiko Gotoh, Alec Ritchie, Hal Broxmeyer

Research output: Contribution to journalArticle

36 Citations (Scopus)

Abstract

Integrin-mediated interaction of hematopoietic progenitor cells with bone marrow stromal extracellular matrix components is important in hematopoiesis. Focal adhesion kinase (pp25FAK)) plays a central role in signal transduction through integrin receptors. We studied matrix-integrin interaction and subsequent signaling in human growth factor-dependent cell line, TF-1. Adherence of unstimulated TF-1 cells to fibronectin-coated wells was blocked by antiintegrin βl and combination of anti-α4 with antiα5 antibodies, indicating α4β1 and α5βl integrin mediated adherence. Steel factor (SLF) increased TF-1 adhesion to fibronectin dose-dependently and 10- 7 mol/L wortmannin suppressed SLF-induced adhesion. Immunoprecipitation and immunoblotting with antiphosphotyrosine antibody showed that adherence of TF1 cells to fibronetin without cytokine caused tyrosine phosphorylation of several proteins identified as pp125(FAK) and paxillin. SLF induced spreading of adherent TF-1 cells and enhanced tyrosine phosphorylation of pp125(FAK) and paxillinin a dose-dependent manner. Treatment with SLF without plating on fibronectin did not induce tyrosine phosphorylation of pp125(FAK). Wortmannin, at 10-7 mol/L, completely abolished SLF-induced enhancement of pp125(FAK) tyrosine phosphorylation, while c-kit autophosphorylation was not affected. This suggests that increase of pp125(FAK) tyrosine phosphorylation was mediated through a wortmannin sensitive pathway, rather than by direct action on c-kit tyrosine kinase. Treatment of adherent TF-1 cells with RGDS peptide plus anti-α4 antibody also inhibited SLF-induced enhancement of pp125(FAK) tyrosine phosphorylation without detachment of TF-1 cells. These data suggest that SLF enhances integrin-fibronectin-dependent tyrosine phosphorylation of pp125(FAK) through activation of integrin ('inside-out' signaling) and following integrin occupancy. This establishes a novel linkage between c-kit/SLF pathway and integrin fibronectin signaling.

Original languageEnglish
Pages (from-to)1574-1584
Number of pages11
JournalBlood
Volume89
Issue number5
StatePublished - 1997

Fingerprint

Paxillin
Focal Adhesion Protein-Tyrosine Kinases
Phosphorylation
Stem Cell Factor
Integrins
Tyrosine
Fibronectins
Antibodies
arginyl-glycyl-aspartyl-serine
Adhesion
Signal transduction
Hematopoiesis
Hematopoietic Stem Cells
Immunoprecipitation
Plating
Immunoblotting
Protein-Tyrosine Kinases
Extracellular Matrix
Anti-Idiotypic Antibodies
Signal Transduction

ASJC Scopus subject areas

  • Hematology

Cite this

Steel factor enhances integrin-mediated tyrosine phosphorylation of focal adhesion kinase (pp125(FAK)) and paxillin. / Takahira, Hiroyuki; Gotoh, Akihiko; Ritchie, Alec; Broxmeyer, Hal.

In: Blood, Vol. 89, No. 5, 1997, p. 1574-1584.

Research output: Contribution to journalArticle

Takahira, Hiroyuki ; Gotoh, Akihiko ; Ritchie, Alec ; Broxmeyer, Hal. / Steel factor enhances integrin-mediated tyrosine phosphorylation of focal adhesion kinase (pp125(FAK)) and paxillin. In: Blood. 1997 ; Vol. 89, No. 5. pp. 1574-1584.
@article{61169eec9bd544a3899cbc9900ac4765,
title = "Steel factor enhances integrin-mediated tyrosine phosphorylation of focal adhesion kinase (pp125(FAK)) and paxillin",
abstract = "Integrin-mediated interaction of hematopoietic progenitor cells with bone marrow stromal extracellular matrix components is important in hematopoiesis. Focal adhesion kinase (pp25FAK)) plays a central role in signal transduction through integrin receptors. We studied matrix-integrin interaction and subsequent signaling in human growth factor-dependent cell line, TF-1. Adherence of unstimulated TF-1 cells to fibronectin-coated wells was blocked by antiintegrin βl and combination of anti-α4 with antiα5 antibodies, indicating α4β1 and α5βl integrin mediated adherence. Steel factor (SLF) increased TF-1 adhesion to fibronectin dose-dependently and 10- 7 mol/L wortmannin suppressed SLF-induced adhesion. Immunoprecipitation and immunoblotting with antiphosphotyrosine antibody showed that adherence of TF1 cells to fibronetin without cytokine caused tyrosine phosphorylation of several proteins identified as pp125(FAK) and paxillin. SLF induced spreading of adherent TF-1 cells and enhanced tyrosine phosphorylation of pp125(FAK) and paxillinin a dose-dependent manner. Treatment with SLF without plating on fibronectin did not induce tyrosine phosphorylation of pp125(FAK). Wortmannin, at 10-7 mol/L, completely abolished SLF-induced enhancement of pp125(FAK) tyrosine phosphorylation, while c-kit autophosphorylation was not affected. This suggests that increase of pp125(FAK) tyrosine phosphorylation was mediated through a wortmannin sensitive pathway, rather than by direct action on c-kit tyrosine kinase. Treatment of adherent TF-1 cells with RGDS peptide plus anti-α4 antibody also inhibited SLF-induced enhancement of pp125(FAK) tyrosine phosphorylation without detachment of TF-1 cells. These data suggest that SLF enhances integrin-fibronectin-dependent tyrosine phosphorylation of pp125(FAK) through activation of integrin ('inside-out' signaling) and following integrin occupancy. This establishes a novel linkage between c-kit/SLF pathway and integrin fibronectin signaling.",
author = "Hiroyuki Takahira and Akihiko Gotoh and Alec Ritchie and Hal Broxmeyer",
year = "1997",
language = "English",
volume = "89",
pages = "1574--1584",
journal = "Blood",
issn = "0006-4971",
publisher = "American Society of Hematology",
number = "5",

}

TY - JOUR

T1 - Steel factor enhances integrin-mediated tyrosine phosphorylation of focal adhesion kinase (pp125(FAK)) and paxillin

AU - Takahira, Hiroyuki

AU - Gotoh, Akihiko

AU - Ritchie, Alec

AU - Broxmeyer, Hal

PY - 1997

Y1 - 1997

N2 - Integrin-mediated interaction of hematopoietic progenitor cells with bone marrow stromal extracellular matrix components is important in hematopoiesis. Focal adhesion kinase (pp25FAK)) plays a central role in signal transduction through integrin receptors. We studied matrix-integrin interaction and subsequent signaling in human growth factor-dependent cell line, TF-1. Adherence of unstimulated TF-1 cells to fibronectin-coated wells was blocked by antiintegrin βl and combination of anti-α4 with antiα5 antibodies, indicating α4β1 and α5βl integrin mediated adherence. Steel factor (SLF) increased TF-1 adhesion to fibronectin dose-dependently and 10- 7 mol/L wortmannin suppressed SLF-induced adhesion. Immunoprecipitation and immunoblotting with antiphosphotyrosine antibody showed that adherence of TF1 cells to fibronetin without cytokine caused tyrosine phosphorylation of several proteins identified as pp125(FAK) and paxillin. SLF induced spreading of adherent TF-1 cells and enhanced tyrosine phosphorylation of pp125(FAK) and paxillinin a dose-dependent manner. Treatment with SLF without plating on fibronectin did not induce tyrosine phosphorylation of pp125(FAK). Wortmannin, at 10-7 mol/L, completely abolished SLF-induced enhancement of pp125(FAK) tyrosine phosphorylation, while c-kit autophosphorylation was not affected. This suggests that increase of pp125(FAK) tyrosine phosphorylation was mediated through a wortmannin sensitive pathway, rather than by direct action on c-kit tyrosine kinase. Treatment of adherent TF-1 cells with RGDS peptide plus anti-α4 antibody also inhibited SLF-induced enhancement of pp125(FAK) tyrosine phosphorylation without detachment of TF-1 cells. These data suggest that SLF enhances integrin-fibronectin-dependent tyrosine phosphorylation of pp125(FAK) through activation of integrin ('inside-out' signaling) and following integrin occupancy. This establishes a novel linkage between c-kit/SLF pathway and integrin fibronectin signaling.

AB - Integrin-mediated interaction of hematopoietic progenitor cells with bone marrow stromal extracellular matrix components is important in hematopoiesis. Focal adhesion kinase (pp25FAK)) plays a central role in signal transduction through integrin receptors. We studied matrix-integrin interaction and subsequent signaling in human growth factor-dependent cell line, TF-1. Adherence of unstimulated TF-1 cells to fibronectin-coated wells was blocked by antiintegrin βl and combination of anti-α4 with antiα5 antibodies, indicating α4β1 and α5βl integrin mediated adherence. Steel factor (SLF) increased TF-1 adhesion to fibronectin dose-dependently and 10- 7 mol/L wortmannin suppressed SLF-induced adhesion. Immunoprecipitation and immunoblotting with antiphosphotyrosine antibody showed that adherence of TF1 cells to fibronetin without cytokine caused tyrosine phosphorylation of several proteins identified as pp125(FAK) and paxillin. SLF induced spreading of adherent TF-1 cells and enhanced tyrosine phosphorylation of pp125(FAK) and paxillinin a dose-dependent manner. Treatment with SLF without plating on fibronectin did not induce tyrosine phosphorylation of pp125(FAK). Wortmannin, at 10-7 mol/L, completely abolished SLF-induced enhancement of pp125(FAK) tyrosine phosphorylation, while c-kit autophosphorylation was not affected. This suggests that increase of pp125(FAK) tyrosine phosphorylation was mediated through a wortmannin sensitive pathway, rather than by direct action on c-kit tyrosine kinase. Treatment of adherent TF-1 cells with RGDS peptide plus anti-α4 antibody also inhibited SLF-induced enhancement of pp125(FAK) tyrosine phosphorylation without detachment of TF-1 cells. These data suggest that SLF enhances integrin-fibronectin-dependent tyrosine phosphorylation of pp125(FAK) through activation of integrin ('inside-out' signaling) and following integrin occupancy. This establishes a novel linkage between c-kit/SLF pathway and integrin fibronectin signaling.

UR - http://www.scopus.com/inward/record.url?scp=0031038001&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0031038001&partnerID=8YFLogxK

M3 - Article

C2 - 9057639

AN - SCOPUS:0031038001

VL - 89

SP - 1574

EP - 1584

JO - Blood

JF - Blood

SN - 0006-4971

IS - 5

ER -