Microtubule-dependent modulation of adhesion complex composition.

{"sci_version": "1.2", "schema": "https://corpus.bntrg.co.uk/sci/1.0/sci.schema.json", "id": "pmid:25526367", "kind": "science-record", "revision": 6, "content_hash": "sha256:64e9bd1811c313b3cb4465c6e1c7b55d19023fd6f02dfa7f32d7ffd968bf0562", "created_at": "2026-11-01T15:40:07Z", "updated_at": "2026-11-01T21:19:56Z", "generator": {"name": "corpus-assay-engine", "run_id": "reading-sci-2026-11-01"}, "paper": {"state": "complete", "title": "Microtubule-dependent modulation of adhesion complex composition.", "doi": "10.1371/journal.pone.0115213", "pmid": "25526367", "authors": ["Daniel H J Ng", "Jonathan D Humphries", "Adam Byron", "Angélique Millon-Frémillon", "Martin J Humphries"], "journal": "PloS one", "year": 2014, "abstract": "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.", "claims": [{"id": "claim-method", "statement": "QSpec [41] was used to identify proteins that were statistically enriched in FN over PDL.", "kind": "methodological", "target": null, "evidence": [{"asset": "source:pmid:25526367", "locator": "sections[14] \"Quantitative MS analysis of isolated ventral plasma membrane complexes upon microtubule disruption\" paragraph 4", "quote": "QSpec [41] was used to identify proteins that were statistically enriched in FN over PDL."}], "state": "complete"}, {"id": "claim-data-access", "statement": "The authors confirm that all data underlying the findings are fully available without restriction. All relevant data are within the paper and its Supporting Information files. 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Here we used label-free quantitative mass spectrometry-based proteomics to determine adhesion compl", "el-free quantitative mass spectrometry-based proteomics to determine adhesion complex changes that o", "lysed either by Western blotting or MS-based proteomics. Initially, serum-starved HFFs adherent to F", "e for future adhesion signaling studies. The proteomics data have been deposited in the ProteomeXcha"], "order_basis": "paper_text_order:496", "id": "assay:1"}, {"role": "data_generation", "definition_id": "ihc", "definition_version": "99e1195c7d6f", "definition_digest": null, "required_operations": [], "support_state": "partial", "source_hashes": {"assay_lexicon": "99e1195c7d6f"}, "confidence": 0.99, "evidence": ["ls of the fibronectin-binding α5β1 integrin. Immunofluorescence analyses confirmed these findings, but revea", "Western blotting (Fig. 3). In agreement with immunofluorescence results, ventral plasma membrane complexes f", "srupt the microtubule network, and fixed and immunostained for actin, vinculin and tubulin (Fig. 2a). I", "glutaraldehyde to preserve microtubules, and immunostained for actin (green), vinculin (red) and tubuli"], "order_basis": "paper_text_order:807", "id": "assay:2"}, {"role": "data_generation", "definition_id": "western_blot", "definition_version": "99e1195c7d6f", "definition_digest": null, "required_operations": [], "support_state": "partial", "source_hashes": {"assay_lexicon": "99e1195c7d6f"}, "confidence": 0.99, "evidence": ["sonication (Fig. 1), and analysed either by Western blotting or MS-based proteomics. Initially, serum-sta", "ehicle (DMSO) or nocodazole, and analysed by Western blotting (Fig. 3). In agreement with immunofluorescen", "plasma membrane complexes were isolated for Western blotting for AC components (α5-integrin, talin, vincu", "d in FN over PDL conditions as identified by Western blotting were found in cluster (iii) (Fig. 3). Unsupe"], "order_basis": "paper_text_order:3226", "id": "assay:3"}, {"role": "analysis", "definition_id": "clustering", "definition_version": "f2132e96ae6c7a4e", "definition_digest": null, "required_operations": ["hierarchical_cluster"], "support_state": "supported", "source_hashes": {"dsl_catalogue": "f2132e96ae6c7a4e"}, "confidence": 0.8, "evidence": ["hierarchical clustering"], "order_basis": "consumes_earlier_output", "id": "assay:4"}]}