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...
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2014-01-01
|
| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC4272306?pdf=render |
| _version_ | 1819212924408823808 |
|---|---|
| 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. |
| first_indexed | 2024-12-23T06:50:42Z |
| format | Article |
| id | doaj.art-ce7c54621b8b4fc593201d8a227423d1 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-23T06:50:42Z |
| publishDate | 2014-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-ce7c54621b8b4fc593201d8a227423d12022-12-21T17:56:26ZengPublic 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.http://europepmc.org/articles/PMC4272306?pdf=render |
| 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 | http://europepmc.org/articles/PMC4272306?pdf=render |
| work_keys_str_mv | AT danielhjng microtubuledependentmodulationofadhesioncomplexcomposition AT jonathandhumphries microtubuledependentmodulationofadhesioncomplexcomposition AT adambyron microtubuledependentmodulationofadhesioncomplexcomposition AT angeliquemillonfremillon microtubuledependentmodulationofadhesioncomplexcomposition AT martinjhumphries microtubuledependentmodulationofadhesioncomplexcomposition |