title: Expanding the Interactome of TES by Exploiting TES Modules with Different Subcellular Localizations.
abstract: The multimodular nature of many eukaryotic proteins underlies their temporal or spatial engagement in a range of protein cocomplexes. Using the multimodule protein testin (TES), we here report a proteomics approach to increase insight in cocomplex diversity. The LIM-domain containing and tumor suppressor protein TES is present at different actin cytoskeleton adhesion structures in cells and influences cell migration, adhesion and spreading. TES module accessibility has been proposed to vary due to conformational switching and variants of TES lacking specific domains target to different subcellular locations. By applying iMixPro AP-MS ("intelligent Mixing of Proteomes"-affinity purification-mass spectrometry) to a set of tagged-TES modular variants, we identified proteins residing in module-specific cocomplexes. The obtained distinct module-specific interactomes combine to a global TES interactome that becomes more extensive and richer in information. Applying pathway analysis to the module interactomes revealed expected actin-related canonical pathways and also less expected pathways. We validated two new TES cocomplex partners: TGFB1I1 and a short form of the glucocorticoid receptor. TES and TGFB1I1 are shown to oppositely affect cell spreading providing biological validity for their copresence in complexes since they act in similar processes.
authors.affiliation: Department of Biochemistry, Ghent University , 9000 Gent, Belgium.
authors.forename: Stefano
authors.identifier: 0000-0003-3675-6849
authors.initials: S
authors.lastname: Sala
authors.affiliation: Department of Biochemistry, Ghent University , 9000 Gent, Belgium.
authors.forename: Marleen
authors.initials: M
authors.lastname: Van Troys
authors.affiliation: Cytoskeleton and Cell Plasticity Lab, Life Sciences Research Unit - FSTC, University of Luxembourg , 4365 Esch-sur-Alzette, Luxembourg.|Laboratory of Experimental Cancer Research, LIH , 1445 Strassen, Luxembourg.
authors.forename: Sandrine
authors.initials: S
authors.lastname: Medves
authors.affiliation: Cytoskeleton and Cell Plasticity Lab, Life Sciences Research Unit - FSTC, University of Luxembourg , 4365 Esch-sur-Alzette, Luxembourg.
authors.forename: Marie
authors.initials: M
authors.lastname: Catillon
authors.affiliation: Department of Biochemistry, Ghent University , 9000 Gent, Belgium.|VIB Medical Biotechnology Center , 9000 Gent, Belgium.
authors.forename: Evy
authors.initials: E
authors.lastname: Timmerman
authors.affiliation: Department of Biochemistry, Ghent University , 9000 Gent, Belgium.|VIB Medical Biotechnology Center , 9000 Gent, Belgium.
authors.forename: An
authors.initials: A
authors.lastname: Staes
authors.affiliation: Cytoskeleton and Cell Plasticity Lab, Life Sciences Research Unit - FSTC, University of Luxembourg , 4365 Esch-sur-Alzette, Luxembourg.
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Schaffner-Reckinger
authors.affiliation: Department of Biochemistry, Ghent University , 9000 Gent, Belgium.|VIB Medical Biotechnology Center , 9000 Gent, Belgium.
authors.forename: Kris
authors.identifier: 0000-0002-4237-0283
authors.initials: K
authors.lastname: Gevaert
authors.affiliation: Department of Biochemistry, Ghent University , 9000 Gent, Belgium.
authors.forename: Christophe
authors.initials: C
authors.lastname: Ampe
mesh_terms: D000199:Actins / Q000378:metabolism; D003598:Cytoskeletal Proteins / Q000378:metabolism*; D022001:Focal Adhesions / Q000378:metabolism; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000378:metabolism; D042541:Intracellular Space / Q000378:metabolism; D060588:LIM Domain Proteins / Q000378:metabolism*; D055503:Protein Multimerization; D040901:Proteomics / Q000379:methods; D016601:RNA-Binding Proteins
chemical_list: D000199:Actins; D003598:Cytoskeletal Proteins; D047908:Intracellular Signaling Peptides and Proteins; D060588:LIM Domain Proteins; D016601:RNA-Binding Proteins; C429688:TES protein, human; C490351:TGFB1I1 protein, human
keywords: VASP; Zyxin; actin; affinity purification−mass spectrometry; cell spreading; focal adhesion; glucocorticoid receptor; pathway analysis; protein−protein interaction; transforming growth factor beta 1 induced 1