title: Polyglutamylation of tubulin as a progressive regulator of in vitro interactions between the microtubule-associated protein Tau and tubulin.
abstract: The multiple functions of microtubules are mediated by various structural and motor microtubule-associated proteins (MAPs). To harmonize these functions in different places of a single cell, the key problem is to regulate the interactions of these proteins with microtubules. The chemical diversity of tubulin isoforms, which constitute the microtubule wall, could represent a molecular basis for this control. Using an in vitro assay of ligand blotting, we found that the microtubule-associated protein Tau interacts differentially with the diverse posttranslationally-modified isotubulins: its binding is mainly restricted to moderately-modified alpha- and beta-tubulin isoforms. We obtained evidence that the recently-discovered polyglutamylation, which consists of the sequential, posttranslational addition of one to six glutamyl units to both alpha- and beta-tubulin subunits, regulates the binding of Tau as a function of its chain length. The relative affinity of Tau, very low for unmodified tubulin, increases progressively for isotubulins carrying from one to three glutamyl units, reaches an optimal value, and then decreases progressively when the polygutamyl chain lengthens up to six residues. Our results suggest that the unmodified C-terminus of tubulin exerts a constitutive inhibition on Tau binding, probably by locking the MAP-binding site, and that this inhibition could be first released and then restored as the polyglutamyl chain grows. As the posttranslational chain does not appear to interact directly with Tau, it is thought that the growth of this chain from one to six glutamyl units causes a progressive, conformational shift in the structure of the C-terminal domain of tubulin, thus leading to the observed modulation of affinity.
authors.affiliation: Laboratoire de Biochimie Cellulaire, Collège de France and Université Pierre et Marie Curie, Paris, France.
authors.forename: D
authors.initials: D
authors.lastname: Boucher
authors.forename: J C
authors.initials: JC
authors.lastname: Larcher
authors.forename: F
authors.initials: F
authors.lastname: Gros
authors.forename: P
authors.initials: P
authors.lastname: Denoulet
mesh_terms: D000818:Animals; D001665:Binding Sites; D001923:Brain Chemistry; D015180:Electrophoresis, Gel, Two-Dimensional; D015151:Immunoblotting; D007526:Isoelectric Point; D051379:Mice; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D008970:Molecular Weight; D011099:Polyglutamic Acid / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D013329:Structure-Activity Relationship; D013381:Subtilisins / Q000378:metabolism; D014404:Tubulin / Q000737:chemistry / Q000378:metabolism*; D016875:tau Proteins / Q000378:metabolism*
chemical_list: D008869:Microtubule-Associated Proteins; D014404:Tubulin; D016875:tau Proteins; D011099:Polyglutamic Acid; D013381:Subtilisins