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PMID: 7537275 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Integrin-dependent translocation of phosphoinositide 3-kinase to the cytoskeleton of thrombin-activated platelets involves specific interactions of p85 alpha with actin filaments and focal adhesion kinase.

The Journal of cell biology ·Vol. 129 ·No. 3 ·1995-05-00 ·Pages 831-42

Guinebault C, Payrastre B, Racaud-Sultan C, Mazarguil H, Breton M, Mauco G, Plantavid M, Chap H

Abstract

Thrombin-induced accumulation of phosphatidylinositol 3,4-bisphosphate (PtdIns(3,4)P2) but not of PtdIns(3,4,5,)P3 is strongly correlated with the relocation to the cytoskeleton of 29% of the p85 alpha regulatory subunit of phosphoinositide 3-kinase (PtdIns 3-kinase) and is accompanied by a significant increase in PtdIns 3-kinase activity in this subcellular fraction. Actually, PtdIns(3,4)P2 accumulation and PtdIns 3-kinase, pp60c-src, and p125FAK translocations as well as aggregation were concomitant events occurring with a distinct lag after actin polymerization. The accumulation of PtdIns(3,4)P2 and the relocalization of PtdIns 3-kinase to the cytoskeleton were both dependent on tyrosine phosphorylation, integrin signaling, and aggregation. Furthermore, although p85 alpha was detected in anti-phosphotyrosine immunoprecipitates obtained from the cytoskeleton of thrombin-activated platelets, we failed to demonstrate tyrosine phosphorylation of cytoskeletal p85 alpha. Tyrphostin treatment clearly reduced its presence in this subcellular fraction, suggesting a physical interaction of p85 alpha with a phosphotyrosyl protein. These data led us to investigate the proteins that are able to interact with PtdIns 3-kinase in the cytoskeleton. We found an association of this enzyme with actin filaments: this interaction was spontaneously restored after one cycle of actin depolymerization-repolymerization in vitro. This association with F-actin appeared to be at least partly indirect, since we demonstrated a thrombin-dependent interaction of p85 alpha with a proline-rich sequence of the tyrosine-phosphorylated cytoskeletal focal adhesion kinase, p125FAK. In addition, we show that PtdIns 3-kinase is significantly activated by the p125FAK proline-rich sequence binding to the src homology 3 domain of p85 alpha subunit. This interaction may represent a new mechanism for PtdIns 3-kinase activation at very specific areas of the cell and indicates that the focal contact-like areas linked to the actin filaments play a critical role in signaling events that occur upon ligand engagement of alpha IIb/beta 3 integrin and platelet aggregation evoked by thrombin.

MeSH Terms
Actin Cytoskeleton/physiology Actins/metabolism Avian Proteins Biological Transport Blood Platelets/drug effects,enzymology Cell Adhesion Molecules/metabolism Cell Compartmentation Cytoskeletal Proteins/metabolism Cytoskeleton/physiology Enzyme Activation Focal Adhesion Kinase 1 Focal Adhesion Protein-Tyrosine Kinases Humans Integrins/metabolism Multienzyme Complexes/metabolism Phosphatidylinositol 3-Kinases Phosphorylation Phosphotransferases (Alcohol Group Acceptor)/metabolism Platelet Activation/physiology Platelet Glycoprotein GPIIb-IIIa Complex Protein-Tyrosine Kinases/metabolism Proto-Oncogene Proteins pp60(c-src)/metabolism Signal Transduction/physiology Thrombin/pharmacology Tyrosine/metabolism
Chemicals
Actins Avian Proteins Cell Adhesion Molecules Cytoskeletal Proteins Integrins Multienzyme Complexes Platelet Glycoprotein GPIIb-IIIa Complex cytoskeletal p85 protein, chicken Tyrosine Phosphatidylinositol 3-Kinases Phosphotransferases (Alcohol Group Acceptor) Protein-Tyrosine Kinases Focal Adhesion Kinase 1 Focal Adhesion Protein-Tyrosine Kinases PTK2 protein, human Proto-Oncogene Proteins pp60(c-src) Thrombin
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Guinebault C
Institut National de la Santé et de la Recherche Médicale, Unité 326, Hôpital Purpan, Toulouse, France.
Payrastre B
Racaud-Sultan C
Mazarguil H
Breton M
Mauco G
Plantavid M
Chap H
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.129.3.831
Published
1995-05-00
Pages
831-42
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2120444
Subset
IM
External Links
PubMed source DOI: 10.1083/jcb.129.3.831 PMC Full Text View journal details
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