Tubulin, actin, and tau protein interactions and the study of their macromolecular assemblies.
The intracellular polymerization of cytoskeletal proteins into their supramolecular assemblies raises many questions regarding the regulatory patterns that control this process. Binding experiments using the ELISA solid phase system, together with protein assembly assays and electron microscopical studies provided clues on the protein-protein associations in the polymerization of tubulin and actin networks. In vitro reconstitution experiments of these cytoskeletal filaments using purified tau, tubulin, and actin proteins were carried out. Tau protein association with tubulin immobilized in a solid phase support system was inhibited by actin monomer, and a higher inhibition was attained in the presence of preassembled actin filaments. Conversely, tubulin and assembled microtubules strongly inhibited tau interaction with actin in the solid phase system. Actin filaments decreased the extent of in vitro tau-induced tubulin assembly. Studies on the morphological aspects of microtubules and actin filaments coexisting in vitro, revealed the association between both cytoskeletal filaments, and in some cases, the presence of fine filamentous structures bridging these polymers. Immunogold studies showed the association of tau along polymerized microtubules and actin filaments, even though a preferential localization of labeled tau with microtubules was revealed. The studies provide further evidence for the involvement of tau protein in modulating the interactions of microtubules and actin polymers in the organization of the cytsokeletal network.
Farias
Gustavo A
GA
Laboratory of Cellular and Molecular Biology, Millennium Institute for Advanced Studies in Cell Biology and Biotechnology (CBB), Universidad de Chile, Las Palmeras 3425, Nuñoa, Santiago, Chile.
Muñoz
Juan P
JP
Garrido
Jorge
J
Maccioni
Ricardo B
RB
D000199:Actins / Q000378:metabolism*; D000818:Animals; D001921:Brain / Q000378:metabolism / Q000648:ultrastructure; D002417:Cattle; D002478:Cells, Cultured; D046911:Macromolecular Substances; D008854:Microscopy, Electron; D008870:Microtubules / Q000502:physiology*; D011108:Polymers / Q000378:metabolism; D011485:Protein Binding; D014404:Tubulin / Q000378:metabolism*; D016875:tau Proteins / Q000378:metabolism*
D000199:Actins; D046911:Macromolecular Substances; D011108:Polymers; D014404:Tubulin; D016875:tau Proteins