MassIVEapplication/xmlftp://massive-ftp.ucsd.edu/x01/MSV000079926/primary200OK1000ProteomicsDr Martin James HumphriesLTQ Orbitrap EliteHomo Sapiens (ncbitaxon:9606)https://massive.ucsd.edu/ProteoSAFe/dataset.jsp?task=4d24b991e54c485a924858d016c54d4aMassIVE266UNIMOD:4 - "Iodoacetamide derivative."UNIMOD:35 - "Oxidation or Hydroxylation."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.Microtubule-dependent modulation of adhesion complex composition.Ng Daniel H J DH, Humphries Jonathan D JD, Byron Adam A, Millon-Frémillon Angélique A, Humphries Martin J MJR 17934, Focal Adhesion, (5-(2-thienylcarbonyl)-1H-benzimidazol-2-yl)-, dlmo, nucleocytoplasm, alphaPS2, DLMO, Rho A, neurotubule, Adherens Junctions, RHO12, hdp-a, dmTAF[[II]]230, dLmo, dLMO, DmelCG1004, Glycoprotein, rho-1, Kinase Inhibitors, symptoms, Inhibitors, DRho, establishment and maintenance of substrate location, alpha2Int, Analysis, Focal Contact, Cell Motility, AAF01186, microtubulus, 8416, Kinase Inhibitor, Mass Spectrum Analysis, nocodazol, Dmrho, increased, Carbamic acid, TFIID TAF250, Analyses, Focal, cel, Cell-Matrix Adherens Junctions, Cold-Insoluble Globulins, DMRHO, Cell Locomotion, nocodazolum, alpha 2-Surface Binding Glycoprotein, 934, free, rhoA, RHO1, actomyosin structure, Rho1, DmelCG8416, Bd, Adhesions, Movements, DmelCG9623, Cold Insoluble Globulins, Dlmo, R-17934, rho1, Adhesion, Locomotion, Actomyosins, Isovinculin, Adherens Junction, microtubuli, Contacts, fliH, associated, Opsonic, Adhesion Plaques, Adhesion Plaque, intracellular, single organism signaling, Plaque, screening, establishment and maintenance of substance location, Motility, alpha Vinculin, dTAF[[II]]230, DmRHO-A, Rho-1, PS2, Fibronectin, Focal Contacts, RHOb, Cell Migration, Cell-Matrix, TAF200, HSC0013, CG1004, TAFII-250, Opsonic Glycoprotein, TAF250/230, Spectrum Analysis, results, Protein Kinase Inhibitor, Dm Rho1, Spectroscopy, Tensin, rhom, (4(S)-trans)-isomer, aPS2, CT27194, single organism localization, TAFII250, RhoN19, Cell-Matrix Adherens Junction, Vinculin (Caenorhabditis elegans clone pRB9.1 protein moiety reduced), (5-(2-thienylcarbonyl)-1H-benzimidazol-2-yl)-carbamic acid methyl ester, RhoA, ve, RHOM, RhoM, internal to cell, RHOHP1, Microtubule, rhoa, Ptd, arha, FN, LETS Proteins, alpha 2 Surface Binding Glycoprotein, D-Rho1, AI326383, nocodazole, Ve, CG8416, Spectrometry, signs, CG9623, RHO, Rho, Junction, CG17603, TAF[[II]], Cell-Matrix Adherens, DMRHOa, Plaques, DMRHOb, NSC238159, Taf250, rhomboid/veinlet, oncodazole, label, SR3-5, Lmo, LMO, ARHD, ARHA, Globulins, hdp, PSalpha2, RHOH12, rho, Protein Kinase, Inhibitor, accessory, TAF230, establishment and maintenance of cellular component location, metavinculin, N-(5-(2-thienoyl)-2-benzimidazolyl)carbamic acid methyl ester, Dmelrho, actomyosin complex, d230, Migration, protein kinase inhibitors, localised, DRORHO, Peptidomics, number, Arhd, Integrin, Arha, dTAFII250, Spectrum Analyses, EfW1, supernumerary, presence, Rho GTPase, ARH12, Opsonic alpha(2)SB Glycoprotein, in, DRhoA, dmTAF1, Taf230, beta-Vinculin, dttg, Mass, establishment and maintenance of position, CG6500, (4(R)-trans)-isomer, If, Mass Spectroscopy, VCL, TAF250, protoplasm, dRhoA, Taf200, beta Vinculin, dTAF[[II]]250, alpha[[PS2]], localized, beadex/dLMO, protoplast, cell, ligand, Y27632, Taf1p, DRho1, integrin, Junctions, Cell Movements, DrhoA, OPN2, dTAF250, Movement, alphaPS2C, RhoHP1, alpha-Vinculin, Drho1, PS 2, dRho1, TAF, methyl N-(5-thenoyl-2-benzimidazolyl)carbamate, DmelCG6500, Peptidomics., Rho kinase, TAF[[II]]250, findings, (trans)-isomer, l(2)52Fa, Proteins, iks, methyl ester, Y-27632, l(3)84Ab, BG:DS00004.13, N-(5-(2-thenoyl)-2-benzimidazolyl)carbamic acid methyl ester, l(2)k02107b, LETS, Cell, NSC-238159, dTAF230, Arha2, methyl (5-(2-thienylcarbonyl))-1H-benzimidazole-2-ylcarbamate, count in organism, MS, Y 27632, p230, Arha1, Protein, TAF[[II]]250/230, TFIID, establishment and maintenance of localization, Cold-Insoluble, dnRho, Mass Spectrum Analyses, Taf[[II]]250, Mass Spectrum, Cell Matrix Adherens Junctions, TAF[[II]]230, increased number, xrhoa, E3.10/J3.8, TAF[II]250, single-organism localization, R17934, inf, NSC 238159, hld, present in greater numbers in organism, dihydrochloride, DmelCG17603, localisation, signalling process, Contact, PS2alpha, alpha[[PS2(ms8)]], Oncodazole, N-(4-pyridyl)-4-(1-aminoethyl)cyclohexanecarboxamide, R17, n(2)k07236, TAF1R 17934, nocodazol, Therapy, treatment, N-(5-(2-thienoyl)-2-benzimidazolyl)carbamic acid methyl ester, (5-(2-thienylcarbonyl)-1H-benzimidazol-2-yl)-, Carbamic acid, nocodazole, methyl ester, nocodazolum, neurotubule, N-(5-(2-thenoyl)-2-benzimidazolyl)carbamic acid methyl ester, 934, R17934, Treatments, NSC-238159, NSC 238159, methyl (5-(2-thienylcarbonyl))-1H-benzimidazole-2-ylcarbamate, NSC238159, disease management., Therapeutic, oncodazole, R-17934, (5-(2-thienylcarbonyl)-1H-benzimidazol-2-yl)-carbamic acid methyl ester, Therapies, Treatment, microtubuli, Oncodazole, R17, Microtubule, microtubulus, methyl N-(5-thenoyl-2-benzimidazolyl)carbamatemicrotubuli, microtubulus., neurotubule, Microtubulemicrotubuli, microtubulus., neurotubule, Microtubule1PXD001183trueProteomic characterisation of microtubule-dependent modulation of adhesion complexesInvestigating the effect of microtubule disruption by nocodazole treatment on isolated adhesion complexes.Tue Jul 19 15:36:00 BST 2016MSV00007992625526367