Tau induces ring and microtubule formation from alphabeta-tubulin dimers under nonassembly conditions.
Tau is a neuronal microtubule-associated protein that plays a central role in many cellular processes, both physiological and pathological, such as axons stabilization and Alzheimer's disease. Despite extensive studies, very little is known about the detailed molecular basis of tau binding to microtubules. We used the four-repeat recombinant htau40 and tubulin dimers to show for the first time that tau is able to induce both microtubule and ring formation from 6S alphabeta tubulin in phosphate buffer without added magnesium (nonassembly conditions). The amount of microtubules or rings formed was protein concentration-, temperature-, and nucleotide-dependent. By means of biophysical approaches, we showed that tau binds to tubulin without global-folding change, detectable by circular dichroism. We also demonstrated that the tau-tubulin interaction follows a ligand-mediated elongation process, with two tau-binding site per tubulin dimer. Moreover, using a tubulin recombinant alpha-tubulin C-terminal fragment (404-451) and a beta-tubulin C-terminal fragment (394-445), we demonstrated the involvement of both of these tubulin regions in tau binding. From this model system, we gain new insight into the mechanisms by which tau binds to tubulin and induces microtubule formation.
Devred
François
F
FRE 2737, ISPDCT, Faculté de Pharmacie, 27 Bd Jean Moulin, 13385 Marseille Cedex 5, France.
Barbier
Pascale
P
Douillard
Soazig
S
Monasterio
Octavio
O
Andreu
José Manuel
JM
Peyrot
Vincent
V
D000818:Animals; D001665:Binding Sites; D001921:Brain / Q000378:metabolism*; D002942:Circular Dichroism; D019281:Dimerization; D005453:Fluorescence; D008274:Magnesium; D008870:Microtubules / Q000378:metabolism*; D008958:Models, Molecular; D009711:Nucleotides / Q000378:metabolism; D010446:Peptide Fragments / Q000378:metabolism; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D012756:Sheep; D013696:Temperature; D014404:Tubulin / Q000737:chemistry / Q000378:metabolism*; D014461:Ultracentrifugation; D016875:tau Proteins / Q000737:chemistry / Q000378:metabolism*
D009711:Nucleotides; D010446:Peptide Fragments; D011994:Recombinant Proteins; D014404:Tubulin; D016875:tau Proteins; D008274:Magnesium