The P2Y2 nucleotide receptor requires interaction with alpha v integrins to access and activate G12.
The P2Y2 nucleotide receptor (P2Y2R) interacts with alpha v integrins to activate G(o) and induce chemotaxis in human 1321N1 astrocytoma cells. In this study, it was determined that the P2Y2R also requires interaction with alpha v integrins to activate G12 and associated signaling pathways that control chemotaxis in 1321N1 cells. Mutation of the Arg-Gly-Asp (RGD) integrin-binding sequence in the first extracellular loop of the human P2Y2R to Arg-Gly-Glu (RGE), which prevents integrin interaction, did not inhibit G(q) or ERK1/2 signaling by the P2Y2R agonist UTP but completely inhibited activation of G12 and G12-mediated events, including Rho activation, cofilin and myosin light chain-2 phosphorylation, stress fiber formation and chemotaxis towards UTP. The involvement of G12 in all these events was verified by using a dominant negative G alpha12 construct. G12 activation by the P2Y2R also was inhibited by anti-alpha v beta5 integrin antibodies and alpha v integrin antisense oligonucleotides, suggesting that alpha v integrin activity and expression are required for the P2Y2R to activate G12. Co-immunoprecipitation experiments confirmed that G alpha12 protein associates with the wild-type P2Y2R and with alpha v integrins but not with the RGE mutant P2Y2R or with alpha3 integrins. Collectively, these results suggest that alpha v integrin complexes provide the P2Y2R with access to G12, thereby allowing activation of this heterotrimeric G protein that controls actin cytoskeletal rearrangements required for chemotaxis.
Liao
Zhongji
Z
Department of Biochemistry, Life Sciences Center, University of Missouri-Columbia, Columbia, MO 65211, USA.
Seye
Cheikh I
CI
Weisman
Gary A
GA
Erb
Laurie
L
D000577:Amides / Q000494:pharmacology; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000494:pharmacology; D024722:Cardiac Myosins / Q000378:metabolism; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects; D051338:Cofilin 1 / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D044365:GTP-Binding Protein alpha Subunits, G12-G13 / Q000235:genetics / Q000378:metabolism*; D043802:GTP-Binding Protein alpha Subunits, Gq-G11 / Q000378:metabolism; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000378:metabolism; D006801:Humans; D039564:Integrin alphaV / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D016023:Integrins / Q000276:immunology / Q000378:metabolism; D009154:Mutation; D018994:Myosin Light Chains / Q000378:metabolism; D016376:Oligonucleotides, Antisense / Q000235:genetics; D037342:Pertussis Toxin / Q000494:pharmacology; D010766:Phosphorylation / Q000187:drug effects; D011485:Protein Binding; D011725:Pyridines / Q000494:pharmacology; D018048:Receptors, Purinergic P2 / Q000235:genetics / Q000378:metabolism*; D058473:Receptors, Purinergic P2Y2; D019030:Receptors, Vitronectin / Q000276:immunology / Q000378:metabolism; D015398:Signal Transduction / Q000235:genetics / Q000502:physiology*; D022502:Stress Fibers / Q000378:metabolism; D014162:Transfection; D014544:Uridine Triphosphate / Q000494:pharmacology; D020741:rho GTP-Binding Proteins / Q000378:metabolism
D000577:Amides; D000911:Antibodies, Monoclonal; C495810:CFL1 protein, human; D051338:Cofilin 1; D004791:Enzyme Inhibitors; D039564:Integrin alphaV; D016023:Integrins; D018994:Myosin Light Chains; D016376:Oligonucleotides, Antisense; C546385:P2RY2 protein, human; D011725:Pyridines; D018048:Receptors, Purinergic P2; D058473:Receptors, Purinergic P2Y2; D019030:Receptors, Vitronectin; C064660:integrin alphaVbeta5; C107184:myosin light chain 2; C108830:Y 27632; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D037342:Pertussis Toxin; D048049:Extracellular Signal-Regulated MAP Kinases; D024722:Cardiac Myosins; D044365:GTP-Binding Protein alpha Subunits, G12-G13; D043802:GTP-Binding Protein alpha Subunits, Gq-G11; D020741:rho GTP-Binding Proteins; D014544:Uridine Triphosphate