title: The conformational state of Tes regulates its zyxin-dependent recruitment to focal adhesions.
abstract: The function of the human Tes protein, which has extensive similarity to zyxin in both sequence and domain organization, is currently unknown. We now show that Tes is a component of focal adhesions that, when expressed, negatively regulates proliferation of T47D breast carcinoma cells. Coimmunoprecipitations demonstrate that in vivo Tes is complexed with actin, Mena, and vasodilator-stimulated phosphoprotein (VASP). Interestingly, the isolated NH2-terminal half of Tes pulls out alpha-actinin and paxillin from cell extracts in addition to actin. The COOH-terminal half recruits zyxin as well as Mena and VASP from cell extracts. These differences suggest that the ability of Tes to associate with alpha-actinin, paxillin, and zyxin is dependent on the conformational state of the molecule. Consistent with this hypothesis, we demonstrate that the two halves of Tes interact with each other in vitro and in vivo. Using fibroblasts lacking Mena and VASP, we show that these proteins are not required to recruit Tes to focal adhesions. However, using RNAi ablation, we demonstrate that zyxin is required to recruit Tes, as well as Mena and VASP, but not vinculin or paxillin, to focal adhesions.
authors.affiliation: European Molecular Biology Laboratory, D-69117 Heidelberg, Germany.
authors.forename: Boyan K
authors.initials: BK
authors.lastname: Garvalov
authors.forename: Theresa E
authors.initials: TE
authors.lastname: Higgins
authors.forename: James D
authors.initials: JD
authors.lastname: Sutherland
authors.forename: Markus
authors.initials: M
authors.lastname: Zettl
authors.forename: Niki
authors.initials: N
authors.lastname: Scaplehorn
authors.forename: Thomas
authors.initials: T
authors.lastname: Köcher
authors.forename: Eugenia
authors.initials: E
authors.lastname: Piddini
authors.forename: Gareth
authors.initials: G
authors.lastname: Griffiths
authors.forename: Michael
authors.initials: M
authors.lastname: Way
mesh_terms: D000185:Actinin / Q000235:genetics / Q000378:metabolism; D001943:Breast Neoplasms / Q000378:metabolism / Q000503:physiopathology; D002277:Carcinoma / Q000378:metabolism; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism; D002448:Cell Adhesion / Q000502:physiology*; D015815:Cell Adhesion Molecules / Q000172:deficiency / Q000235:genetics; D002455:Cell Division / Q000235:genetics; D003598:Cytoskeletal Proteins / Q000235:genetics / Q000378:metabolism; D005057:Eukaryotic Cells / Q000378:metabolism* / Q000648:ultrastructure; D005260:Female; D022001:Focal Adhesions / Q000378:metabolism* / Q000648:ultrastructure; D016147:Genes, Tumor Suppressor; D006023:Glycoproteins; D049452:Green Fluorescent Proteins; D006367:HeLa Cells; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D060588:LIM Domain Proteins; D008164:Luminescent Proteins; D008667:Metalloproteins / Q000235:genetics / Q000378:metabolism*; D008840:Microfilament Proteins; D008968:Molecular Conformation; D051419:Paxillin; D010750:Phosphoproteins / Q000172:deficiency / Q000235:genetics / Q000378:metabolism; D017434:Protein Structure, Tertiary / Q000235:genetics; D021381:Protein Transport / Q000235:genetics; D016601:RNA-Binding Proteins; D011993:Recombinant Fusion Proteins; D022502:Stress Fibers / Q000235:genetics / Q000378:metabolism; D025521:Tumor Suppressor Proteins / Q000235:genetics / Q000378:metabolism*; D016596:Vinculin / Q000235:genetics / Q000378:metabolism; D060589:Zyxin; D000098662:Vasodilator-Stimulated Phosphoprotein
chemical_list: D002352:Carrier Proteins; D015815:Cell Adhesion Molecules; D003598:Cytoskeletal Proteins; C102176:Enah protein, mouse; D006023:Glycoproteins; D018398:Homeodomain Proteins; D060588:LIM Domain Proteins; D008164:Luminescent Proteins; D008667:Metalloproteins; D008840:Microfilament Proteins; C496061:PXN protein, human; D051419:Paxillin; D010750:Phosphoproteins; D016601:RNA-Binding Proteins; D011993:Recombinant Fusion Proteins; C429688:TES protein, human; D025521:Tumor Suppressor Proteins; C092842:ZYX protein, human; C498285:Zyx protein, mouse; D060589:Zyxin; D000098662:Vasodilator-Stimulated Phosphoprotein; D000185:Actinin; D016596:Vinculin; D049452:Green Fluorescent Proteins