Microtubule-dependent modulation of adhesion complex composition.
The microtubule network regulates the turnover of integrin-containing adhesion complexes to stimulate cell migration. Disruption of the microtubule network results in an enlargement of adhesion complex size due to increased RhoA-stimulated actomyosin contractility, and inhibition of adhesion complex...
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Public Library of Science (PLoS)
2014-01-01
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| Series: | PLoS ONE |
| Online Access: | https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0115213&type=printable |
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| author | Daniel H J Ng Jonathan D Humphries Adam Byron Angélique Millon-Frémillon Martin J Humphries |
| author_facet | Daniel H J Ng Jonathan D Humphries Adam Byron Angélique Millon-Frémillon Martin J Humphries |
| author_sort | Daniel H J Ng |
| collection | DOAJ |
| description | The microtubule network regulates the turnover of integrin-containing adhesion complexes to stimulate cell migration. Disruption of the microtubule network results in an enlargement of adhesion complex size due to increased RhoA-stimulated actomyosin contractility, and inhibition of adhesion complex turnover; however, the microtubule-dependent changes in adhesion complex composition have not been studied in a global, unbiased manner. Here we used label-free quantitative mass spectrometry-based proteomics to determine adhesion complex changes that occur upon microtubule disruption with nocodazole. Nocodazole-treated cells displayed an increased abundance of the majority of known adhesion complex components, but no change in the levels of the fibronectin-binding α5β1 integrin. Immunofluorescence analyses confirmed these findings, but revealed a change in localisation of adhesion complex components. Specifically, in untreated cells, α5-integrin co-localised with vinculin at peripherally located focal adhesions and with tensin at centrally located fibrillar adhesions. In nocodazole-treated cells, however, α5-integrin was found in both peripherally located and centrally located adhesion complexes that contained both vinculin and tensin, suggesting a switch in the maturation state of adhesion complexes to favour focal adhesions. Moreover, the switch to focal adhesions was confirmed to be force-dependent as inhibition of cell contractility with the Rho-associated protein kinase inhibitor, Y-27632, prevented the nocodazole-induced conversion. These results highlight a complex interplay between the microtubule cytoskeleton, adhesion complex maturation state and intracellular contractile force, and provide a resource for future adhesion signaling studies. The proteomics data have been deposited in the ProteomeXchange with identifier PXD001183. |
| format | Article |
| id | doaj-art-ce7c54621b8b4fc593201d8a227423d1 |
| institution | OA Journals |
| issn | 1932-6203 |
| language | English |
| publishDate | 2014-01-01 |
| publisher | Public Library of Science (PLoS) |
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| series | PLoS ONE |
| spelling | doaj-art-ce7c54621b8b4fc593201d8a227423d12025-08-20T02:15:15ZengPublic Library of Science (PLoS)PLoS ONE1932-62032014-01-01912e11521310.1371/journal.pone.0115213Microtubule-dependent modulation of adhesion complex composition.Daniel H J NgJonathan D HumphriesAdam ByronAngélique Millon-FrémillonMartin J HumphriesThe microtubule network regulates the turnover of integrin-containing adhesion complexes to stimulate cell migration. Disruption of the microtubule network results in an enlargement of adhesion complex size due to increased RhoA-stimulated actomyosin contractility, and inhibition of adhesion complex turnover; however, the microtubule-dependent changes in adhesion complex composition have not been studied in a global, unbiased manner. Here we used label-free quantitative mass spectrometry-based proteomics to determine adhesion complex changes that occur upon microtubule disruption with nocodazole. Nocodazole-treated cells displayed an increased abundance of the majority of known adhesion complex components, but no change in the levels of the fibronectin-binding α5β1 integrin. Immunofluorescence analyses confirmed these findings, but revealed a change in localisation of adhesion complex components. Specifically, in untreated cells, α5-integrin co-localised with vinculin at peripherally located focal adhesions and with tensin at centrally located fibrillar adhesions. In nocodazole-treated cells, however, α5-integrin was found in both peripherally located and centrally located adhesion complexes that contained both vinculin and tensin, suggesting a switch in the maturation state of adhesion complexes to favour focal adhesions. Moreover, the switch to focal adhesions was confirmed to be force-dependent as inhibition of cell contractility with the Rho-associated protein kinase inhibitor, Y-27632, prevented the nocodazole-induced conversion. These results highlight a complex interplay between the microtubule cytoskeleton, adhesion complex maturation state and intracellular contractile force, and provide a resource for future adhesion signaling studies. The proteomics data have been deposited in the ProteomeXchange with identifier PXD001183.https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0115213&type=printable |
| spellingShingle | Daniel H J Ng Jonathan D Humphries Adam Byron Angélique Millon-Frémillon Martin J Humphries Microtubule-dependent modulation of adhesion complex composition. PLoS ONE |
| title | Microtubule-dependent modulation of adhesion complex composition. |
| title_full | Microtubule-dependent modulation of adhesion complex composition. |
| title_fullStr | Microtubule-dependent modulation of adhesion complex composition. |
| title_full_unstemmed | Microtubule-dependent modulation of adhesion complex composition. |
| title_short | Microtubule-dependent modulation of adhesion complex composition. |
| title_sort | microtubule dependent modulation of adhesion complex composition |
| url | https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0115213&type=printable |
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