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Syndecan-4/PAR-3 signaling regulates focal adhesion dynamics in mesenchymal cells
BACKGROUND: Syndecans regulate cell migration thus having key roles in scarring and wound healing processes. Our previous results have shown that Thy-1/CD90 can engage both αvβ3 integrin and Syndecan-4 expressed on the surface of astrocytes to induce cell migration. Despite a well-described role of...
Autores principales: | , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7433185/ https://www.ncbi.nlm.nih.gov/pubmed/32811537 http://dx.doi.org/10.1186/s12964-020-00629-3 |
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author | Valdivia, Alejandra Cárdenas, Areli Brenet, Marianne Maldonado, Horacio Kong, Milene Díaz, Jorge Burridge, Keith Schneider, Pascal San Martín, Alejandra García-Mata, Rafael Quest, Andrew F. G. Leyton, Lisette |
author_facet | Valdivia, Alejandra Cárdenas, Areli Brenet, Marianne Maldonado, Horacio Kong, Milene Díaz, Jorge Burridge, Keith Schneider, Pascal San Martín, Alejandra García-Mata, Rafael Quest, Andrew F. G. Leyton, Lisette |
author_sort | Valdivia, Alejandra |
collection | PubMed |
description | BACKGROUND: Syndecans regulate cell migration thus having key roles in scarring and wound healing processes. Our previous results have shown that Thy-1/CD90 can engage both αvβ3 integrin and Syndecan-4 expressed on the surface of astrocytes to induce cell migration. Despite a well-described role of Syndecan-4 during cell movement, information is scarce regarding specific Syndecan-4 partners involved in Thy-1/CD90-stimulated cell migration. METHODS: Mass spectrometry (MS) analysis of complexes precipitated with the Syndecan-4 cytoplasmic tail peptide was used to identify potential Syndecan-4-binding partners. The interactions found by MS were validated by immunoprecipitation and proximity ligation assays. The conducted research employed an array of genetic, biochemical and pharmacological approaches, including: PAR-3, Syndecan-4 and Tiam1 silencing, active Rac1 GEFs affinity precipitation, and video microscopy. RESULTS: We identified PAR-3 as a Syndecan-4-binding protein. Its interaction depended on the carboxy-terminal EFYA sequence present on Syndecan-4. In astrocytes where PAR-3 expression was reduced, Thy-1-induced cell migration and focal adhesion disassembly was impaired. This effect was associated with a sustained Focal Adhesion Kinase activation in the siRNA-PAR-3 treated cells. Our data also show that Thy-1/CD90 activates Tiam1, a PAR-3 effector. Additionally, we found that after Syndecan-4 silencing, Tiam1 activation was decreased and it was no longer recruited to the membrane. Syndecan-4/PAR-3 interaction and the alteration in focal adhesion dynamics were validated in mouse embryonic fibroblast (MEF) cells, thereby identifying this novel Syndecan-4/PAR-3 signaling complex as a general mechanism for mesenchymal cell migration involved in Thy-1/CD90 stimulation. CONCLUSIONS: The newly identified Syndecan-4/PAR-3 signaling complex participates in Thy-1/CD90-induced focal adhesion disassembly in mesenchymal cells. The mechanism involves focal adhesion kinase dephosphorylation and Tiam1 activation downstream of Syndecan-4/PAR-3 signaling complex formation. Additionally, PAR-3 is defined here as a novel adhesome-associated component with an essential role in focal adhesion disassembly during polarized cell migration. These novel findings uncover signaling mechanisms regulating cell migration, thereby opening up new avenues for future research on Syndecan-4/PAR-3 signaling in processes such as wound healing and scarring. GRAPHICAL ABSTRACT: [Image: see text] |
format | Online Article Text |
id | pubmed-7433185 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-74331852020-08-19 Syndecan-4/PAR-3 signaling regulates focal adhesion dynamics in mesenchymal cells Valdivia, Alejandra Cárdenas, Areli Brenet, Marianne Maldonado, Horacio Kong, Milene Díaz, Jorge Burridge, Keith Schneider, Pascal San Martín, Alejandra García-Mata, Rafael Quest, Andrew F. G. Leyton, Lisette Cell Commun Signal Research BACKGROUND: Syndecans regulate cell migration thus having key roles in scarring and wound healing processes. Our previous results have shown that Thy-1/CD90 can engage both αvβ3 integrin and Syndecan-4 expressed on the surface of astrocytes to induce cell migration. Despite a well-described role of Syndecan-4 during cell movement, information is scarce regarding specific Syndecan-4 partners involved in Thy-1/CD90-stimulated cell migration. METHODS: Mass spectrometry (MS) analysis of complexes precipitated with the Syndecan-4 cytoplasmic tail peptide was used to identify potential Syndecan-4-binding partners. The interactions found by MS were validated by immunoprecipitation and proximity ligation assays. The conducted research employed an array of genetic, biochemical and pharmacological approaches, including: PAR-3, Syndecan-4 and Tiam1 silencing, active Rac1 GEFs affinity precipitation, and video microscopy. RESULTS: We identified PAR-3 as a Syndecan-4-binding protein. Its interaction depended on the carboxy-terminal EFYA sequence present on Syndecan-4. In astrocytes where PAR-3 expression was reduced, Thy-1-induced cell migration and focal adhesion disassembly was impaired. This effect was associated with a sustained Focal Adhesion Kinase activation in the siRNA-PAR-3 treated cells. Our data also show that Thy-1/CD90 activates Tiam1, a PAR-3 effector. Additionally, we found that after Syndecan-4 silencing, Tiam1 activation was decreased and it was no longer recruited to the membrane. Syndecan-4/PAR-3 interaction and the alteration in focal adhesion dynamics were validated in mouse embryonic fibroblast (MEF) cells, thereby identifying this novel Syndecan-4/PAR-3 signaling complex as a general mechanism for mesenchymal cell migration involved in Thy-1/CD90 stimulation. CONCLUSIONS: The newly identified Syndecan-4/PAR-3 signaling complex participates in Thy-1/CD90-induced focal adhesion disassembly in mesenchymal cells. The mechanism involves focal adhesion kinase dephosphorylation and Tiam1 activation downstream of Syndecan-4/PAR-3 signaling complex formation. Additionally, PAR-3 is defined here as a novel adhesome-associated component with an essential role in focal adhesion disassembly during polarized cell migration. These novel findings uncover signaling mechanisms regulating cell migration, thereby opening up new avenues for future research on Syndecan-4/PAR-3 signaling in processes such as wound healing and scarring. GRAPHICAL ABSTRACT: [Image: see text] BioMed Central 2020-08-18 /pmc/articles/PMC7433185/ /pubmed/32811537 http://dx.doi.org/10.1186/s12964-020-00629-3 Text en © The Author(s) 2020 Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data. |
spellingShingle | Research Valdivia, Alejandra Cárdenas, Areli Brenet, Marianne Maldonado, Horacio Kong, Milene Díaz, Jorge Burridge, Keith Schneider, Pascal San Martín, Alejandra García-Mata, Rafael Quest, Andrew F. G. Leyton, Lisette Syndecan-4/PAR-3 signaling regulates focal adhesion dynamics in mesenchymal cells |
title | Syndecan-4/PAR-3 signaling regulates focal adhesion dynamics in mesenchymal cells |
title_full | Syndecan-4/PAR-3 signaling regulates focal adhesion dynamics in mesenchymal cells |
title_fullStr | Syndecan-4/PAR-3 signaling regulates focal adhesion dynamics in mesenchymal cells |
title_full_unstemmed | Syndecan-4/PAR-3 signaling regulates focal adhesion dynamics in mesenchymal cells |
title_short | Syndecan-4/PAR-3 signaling regulates focal adhesion dynamics in mesenchymal cells |
title_sort | syndecan-4/par-3 signaling regulates focal adhesion dynamics in mesenchymal cells |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7433185/ https://www.ncbi.nlm.nih.gov/pubmed/32811537 http://dx.doi.org/10.1186/s12964-020-00629-3 |
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