Kinetic stabilization of microtubule dynamics at steady state by tau and microtubule-binding domains of tau.
Tau is a neuronal microtubule-associated protein that plays an important role in stabilizing axonal microtubules and maintaining neuronal processes. To investigate the mechanisms by which tau performs these functions, we have determined the actions of full-length adult tau and tau peptides corresponding to two different microtubule-binding domains of tau (the first repeat, R1, VRSKIGSTENLKHQPGGG, and the first interrepeat, R1-R2 IR, KVQIINKK) on the growing and shortening dynamics at the plus ends of individual microtubules at steady state. Tau suppressed steady-state microtubule dynamics at very low molar ratios of tau to tubulin. At the lowest ratios examined (tau:tubulin ratios of 1:175 and 1:85), suppression of dynamics occurred in the absence of a detectable change in polymer mass. Tau reduced the mean rate and extent of shortening and, in contrast to previous work carried out under conditions of net polymer gain, tau also suppressed the mean rate and extent of growing. Tau also strongly increased the rescue frequency, it moderately suppressed the catastrophe frequency and it strongly increased the percentage of total time that the microtubules spent in an attenuated (pause) state, neither growing nor shortening detectably. In addition, both the R1 and R1-R2 IR tau peptides suppressed steady-state microtubule dynamics in a sequence-specific manner and in a manner that was qualitatively indistinguishable from full-length tau. The data provide significant support for a mechanism in which the binding of tau to individual tubulin subunits in microtubules induces a conformational change that strengthens inter-tubulin bonding.
Panda
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Department of Biological Sciences, University of California, Santa Barbara 93106, USA.
Goode
B L
BL
Feinstein
S C
SC
Wilson
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L
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D001923:Brain Chemistry; D002417:Cattle; D004355:Drug Stability; D066298:In Vitro Techniques; D007700:Kinetics; D008870:Microtubules / Q000737:chemistry / Q000378:metabolism*; D008954:Models, Biological; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011108:Polymers / Q000737:chemistry / Q000378:metabolism; D011487:Protein Conformation; D051381:Rats; D013816:Thermodynamics; D014404:Tubulin / Q000737:chemistry / Q000378:metabolism; D016875:tau Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
D010446:Peptide Fragments; D011108:Polymers; D014404:Tubulin; D016875:tau Proteins