Osteopontin activation of c-src in human melanoma cells requires the cytoplasmic domain of the integrin alpha v-subunit

M Chellaiah; C Fitzgerald; E J Filardo; D A Cheresh; K A Hruska

doi:10.1210/endo.137.6.8641196

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Osteopontin activation of c-src in human melanoma cells requires the cytoplasmic domain of the integrin alpha v-subunit

Journal article

Open access

Peer reviewed

Osteopontin activation of c-src in human melanoma cells requires the cytoplasmic domain of the integrin alpha v-subunit

M Chellaiah, C Fitzgerald, E J Filardo, D A Cheresh and K A Hruska

Endocrinology (Philadelphia), Vol.137(6), pp.2432-2440

06/1996

DOI: 10.1210/endo.137.6.8641196

PMID: 8641196

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Abstract

In human melanoma cells, expression of the alpha v beta 3 integrin is correlated with the metastatic potential. The expression of osteopontin (OPN or OP), a protein ligand for the integrin alpha v beta 3, also correlates with metastatic potential of some tumors. Analysis of signal transduction, stimulated by OPN/alpha v beta 3 in human melanoma cells (M21), revealed activation of pp60c-src associated with the integrin. pp60c-src stimulation by OPN was dose dependent, and it was inhibited in vitro by a tyrosine kinase inhibitor, herbimycin-A. To determine the need for the cytoplasmic domain of the alpha v-subunit, in the association of pp60c-src with alpha v beta 3, a cell line expressing truncated alpha v was studied. M21-L cells lacked alpha v expression but stably transfected with complementary DNAs encoding alpha v full length protein alpha v 1018 or alpha v 995 (lacking 23 carboxyl-terminal amino acids), and a fibroblast cell line (FG) expressing alpha v beta 5 but not alpha v beta 3, were used. Western analysis and immune complex kinase assays of anti- alpha v immunoprecipitates demonstrated that M21-L/alpha v995 cells did not exhibit pp60c-src association with alpha v, whereas the alpha v1018 complementary DNA transfected cells and FG cells had pp60c-src associated with the alpha v integrins. Immunofluorescence analysis revealed pp60c-src, alpha v beta 3 integrin, and actin distribution along the plasma membrane of M21 cells. 35S-labeling of cells and analysis of complexes immunoprecipitated by a monoclonal antibody against alpha v beta 3 demonstrated association of actin with the immune complexes. We conclude that OPN stimulates pp60c-src kinase activity associated with the alpha v beta 3 integrin and that the association requires the cytoplasmic tail of the alpha v chain.

Phosphorylation

Melanoma - metabolism

Amino Acid Sequence

Integrin alpha5

Recombinant Fusion Proteins - pharmacology

Humans

Molecular Sequence Data

Osteopontin

Enzyme Activation - drug effects

Cell Membrane - chemistry

Immunosorbent Techniques

Receptors, Vitronectin - analysis

Sialoglycoproteins - pharmacology

Actins - analysis

Proto-Oncogene Proteins pp60(c-src) - metabolism

Fluorescent Antibody Technique

Antigens, CD - physiology

Antigens, CD - chemistry

Receptors, Vitronectin - metabolism

Tumor Cells, Cultured

Proto-Oncogene Proteins pp60(c-src) - analysis

Details

Title: Subtitle: Osteopontin activation of c-src in human melanoma cells requires the cytoplasmic domain of the integrin alpha v-subunit
Creators: M Chellaiah - Renal Division, Jewish Hospital of St. Louis, Missouri 63110, USA
C Fitzgerald
E J Filardo
D A Cheresh
K A Hruska
Resource Type: Journal article
Publication Details: Endocrinology (Philadelphia), Vol.137(6), pp.2432-2440
Publisher: United States
DOI: 10.1210/endo.137.6.8641196
PMID: 8641196
ISSN: 0013-7227
eISSN: 1945-7170
Grant note: R01AR-41677 / NIAMS NIH HHS T32DK-07679 / NIDDK NIH HHS
Language: English
Date published: 06/1996
Academic Unit: Pharmaceutical Sciences and Experimental Therapeutics; Surgery; Internal Medicine
Record Identifier: 9984051581102771

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