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Mechanisms and consequences of agonist-induced talin recruitment to platelet integrin αIIbβ3

Platelet aggregation requires agonist-induced αIIbβ3 activation, a process mediated by Rap1 and talin. To study mechanisms, we engineered αIIbβ3 Chinese hamster ovary (CHO) cells to conditionally express talin and protease-activated receptor (PAR) thrombin receptors. Human PAR1 or murine PAR4 stimul...

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Autores principales: Watanabe, Naohide, Bodin, Laurent, Pandey, Manjula, Krause, Matthias, Coughlin, Shaun, Boussiotis, Vassiliki A., Ginsberg, Mark H., Shattil, Sanford J.
Formato: Texto
Lenguaje:English
Publicado: The Rockefeller University Press 2008
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2442211/
https://www.ncbi.nlm.nih.gov/pubmed/18573917
http://dx.doi.org/10.1083/jcb.200803094
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author Watanabe, Naohide
Bodin, Laurent
Pandey, Manjula
Krause, Matthias
Coughlin, Shaun
Boussiotis, Vassiliki A.
Ginsberg, Mark H.
Shattil, Sanford J.
author_facet Watanabe, Naohide
Bodin, Laurent
Pandey, Manjula
Krause, Matthias
Coughlin, Shaun
Boussiotis, Vassiliki A.
Ginsberg, Mark H.
Shattil, Sanford J.
author_sort Watanabe, Naohide
collection PubMed
description Platelet aggregation requires agonist-induced αIIbβ3 activation, a process mediated by Rap1 and talin. To study mechanisms, we engineered αIIbβ3 Chinese hamster ovary (CHO) cells to conditionally express talin and protease-activated receptor (PAR) thrombin receptors. Human PAR1 or murine PAR4 stimulation activates αIIbβ3, which was measured with antibody PAC-1, indicating complete pathway reconstitution. Knockdown of Rap1–guanosine triphosphate–interacting adaptor molecule (RIAM), a Rap1 effector, blocks this response. In living cells, RIAM overexpression stimulates and RIAM knockdown blocks talin recruitment to αIIbβ3, which is monitored by bimolecular fluorescence complementation. Mutations in talin or β3 that disrupt their mutual interaction block both talin recruitment and αIIbβ3 activation. However, one talin mutant (L325R) is recruited to αIIbβ3 but cannot activate it. In platelets, RIAM localizes to filopodia and lamellipodia, and, in megakaryocytes, RIAM knockdown blocks PAR4-mediated αIIbβ3 activation. The RIAM-related protein lamellipodin promotes talin recruitment and αIIbβ3 activity in CHO cells but is not expressed in megakaryocytes or platelets. Thus, talin recruitment to αIIbβ3 by RIAM mediates agonist-induced αIIbβ3 activation, with implications for hemostasis and thrombosis.
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spelling pubmed-24422112008-12-30 Mechanisms and consequences of agonist-induced talin recruitment to platelet integrin αIIbβ3 Watanabe, Naohide Bodin, Laurent Pandey, Manjula Krause, Matthias Coughlin, Shaun Boussiotis, Vassiliki A. Ginsberg, Mark H. Shattil, Sanford J. J Cell Biol Research Articles Platelet aggregation requires agonist-induced αIIbβ3 activation, a process mediated by Rap1 and talin. To study mechanisms, we engineered αIIbβ3 Chinese hamster ovary (CHO) cells to conditionally express talin and protease-activated receptor (PAR) thrombin receptors. Human PAR1 or murine PAR4 stimulation activates αIIbβ3, which was measured with antibody PAC-1, indicating complete pathway reconstitution. Knockdown of Rap1–guanosine triphosphate–interacting adaptor molecule (RIAM), a Rap1 effector, blocks this response. In living cells, RIAM overexpression stimulates and RIAM knockdown blocks talin recruitment to αIIbβ3, which is monitored by bimolecular fluorescence complementation. Mutations in talin or β3 that disrupt their mutual interaction block both talin recruitment and αIIbβ3 activation. However, one talin mutant (L325R) is recruited to αIIbβ3 but cannot activate it. In platelets, RIAM localizes to filopodia and lamellipodia, and, in megakaryocytes, RIAM knockdown blocks PAR4-mediated αIIbβ3 activation. The RIAM-related protein lamellipodin promotes talin recruitment and αIIbβ3 activity in CHO cells but is not expressed in megakaryocytes or platelets. Thus, talin recruitment to αIIbβ3 by RIAM mediates agonist-induced αIIbβ3 activation, with implications for hemostasis and thrombosis. The Rockefeller University Press 2008-06-30 /pmc/articles/PMC2442211/ /pubmed/18573917 http://dx.doi.org/10.1083/jcb.200803094 Text en © 2008 Watanabe et al. This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.jcb.org/misc/terms.shtml). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/).
spellingShingle Research Articles
Watanabe, Naohide
Bodin, Laurent
Pandey, Manjula
Krause, Matthias
Coughlin, Shaun
Boussiotis, Vassiliki A.
Ginsberg, Mark H.
Shattil, Sanford J.
Mechanisms and consequences of agonist-induced talin recruitment to platelet integrin αIIbβ3
title Mechanisms and consequences of agonist-induced talin recruitment to platelet integrin αIIbβ3
title_full Mechanisms and consequences of agonist-induced talin recruitment to platelet integrin αIIbβ3
title_fullStr Mechanisms and consequences of agonist-induced talin recruitment to platelet integrin αIIbβ3
title_full_unstemmed Mechanisms and consequences of agonist-induced talin recruitment to platelet integrin αIIbβ3
title_short Mechanisms and consequences of agonist-induced talin recruitment to platelet integrin αIIbβ3
title_sort mechanisms and consequences of agonist-induced talin recruitment to platelet integrin αiibβ3
topic Research Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2442211/
https://www.ncbi.nlm.nih.gov/pubmed/18573917
http://dx.doi.org/10.1083/jcb.200803094
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