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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...
Autores principales: | , , , , , , , |
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Formato: | Texto |
Lenguaje: | English |
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The Rockefeller University Press
2008
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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. |
format | Text |
id | pubmed-2442211 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2008 |
publisher | The Rockefeller University Press |
record_format | MEDLINE/PubMed |
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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