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Interaction Between the P2Y2 Nucleotide Receptor and Filamin A Regulates Cell Motility
Author(s) -
Yu Ningpu,
Weisman Gary A,
Seye Cheikh I
Publication year - 2006
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.20.4.a114-b
Subject(s) - filamin , flna , microbiology and biotechnology , actin cytoskeleton , focal adhesion , motility , biology , cytoskeleton , actin , actin binding protein , cell migration , integrin , receptor , signal transduction , cell , biochemistry
Migration of vascular smooth muscle cells (SMC) is an important feature of atherosclerosis. Focal adhesions and actin cytoskeleton are involved in cell growth, shape, and movement. Proteins that target the polymerization of actin filaments and link integrins to the actin cytoskeleton play an important role in cell motility. Many connections between integrins and the actin cytoskeleton are mediated by actin binding proteins. We have reported that activation of the P2Y 2 nucleotide receptor (P2Y 2 R) mediates intimal hyperplasia in collared rabbit arteries. Here, we identify filamin A (FLNA) as a P2Y 2 R receptor‐binding protein using the yeast two‐hybrid assay. UTP, a preferential agonist of the P2Y 2 R, causes actin cytoskeletal reorganization in human coronary artery SMC (CASMC). We also found that P2Y 2 R activation leads to FLNA phosphorylation, its dissociation from β 1 ‐integrin, and focal adhesion formation in CASMC. UTP‐treated CASMC also showed increased cell motility in a chemotaxis assay. To determine the molecular determinants of FLNA binding to the P2Y 2 R, we mapped the FLNA binding site in the P2Y 2 R and deleted amino acids in the C‐terminal domain of the receptor that produced a mutant P2Y 2 R defective in FLNA binding. We showed that expression of this P2Y 2 R mutant receptor suppressed UTP‐induced focal adhesion formation and cell motility. This study clearly identified the molecular mechanism whereby FLNA binding to the P2Y 2 R mediates cell migration and suggests a potential target for the pharmacological control of vascular disease.