New insights into tau–microtubules interaction revealed by isothermal titration calorimetry

Tsvetkov, Philipp O.; Makarov, Alexander А.; Malesinski, Soazig; Peyrot, Vincent; Devred, François

doi:10.1016/j.biochi.2011.09.011

Cited by 25 publications

(22 citation statements)

References 19 publications

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“…Moreover, we characterized two classes of hotspots based on their FRET intensities. We recently demonstrated using isothermal titration calorimetry that Tau binds to tubulin at a high affinity binding site with a stoichiometry of 0.2 and at a low affinity binding site with a stoichiometry of 0.8 (Tsvetkov et al, 2012). These two classes of binding sites might be associated with the high and low FRET hotspots found in the present study.…”

Section: Discussionsupporting

confidence: 60%

Molecular Mechanisms of Tau Binding to Microtubule and its Role in Microtubule Dynamics in Live Cells

Breuzard

Hubert

Nouar

et al. 2013

Journal of Cell Science

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SummaryDespite extensive studies, the molecular mechanisms of Tau binding to microtubules (MTs) and its consequences on MT stability still remain unclear. It is especially true in cells where the spatiotemporal distribution of Tau-MT interactions is unknown. Using Förster resonance energy transfer (FRET), we showed that the Tau-MT interaction was distributed along MTs in periodic hotspots of high and low FRET intensities. Fluorescence recovery after photobleaching (FRAP) revealed a two-phase exchange of Tau with MTs as a rapid diffusion followed by a slower binding phase. A real-time FRET assay showed that high FRET occurred simultaneously with rescue and pause transitions at MT ends. To further explore the functional interaction of Tau with MTs, the binding of paclitaxel (PTX), tubulin acetylation induced by trichostatin A (TSA), and the expression of non-acetylatable tubulin were used. With PTX and TSA, FRAP curves best fitted a single phase with a long time constant, whereas with non-acetylatable a-tubulin, curves best fitted a two phase recovery. Upon incubation with PTX and TSA, the number of high and low FRET hotspots decreased by up to 50% and no hotspot was observed during rescue and pause transitions. In the presence of non-acetylatable a-tubulin, a 34% increase in low FRET hotspots occurred, and our real-time FRET assay revealed that low FRET hotspots appeared with MTs recovering growth. In conclusion, we have identified, by FRET and FRAP, a discrete Tau-MT interaction, in which Tau could induce conformational changes of MTs, favoring recovery of MT self-assembly.

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Section: Discussionsupporting

confidence: 60%

Molecular Mechanisms of Tau Binding to Microtubule and its Role in Microtubule Dynamics in Live Cells

Breuzard

Hubert

Nouar

et al. 2013

Journal of Cell Science

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“…In healthy cells, these proteins are soluble and do not exhibit pathogenic properties. They usually play an important role in different cell processes: SOD1 is implicated in oxidative reactions, FUS and TDP-43 in RNA metabolism, and Tau protein regulates microtubule dynamics by interacting with tubulin 37 . Different endogenous factors such as mutations, hyper phosphorylation, isomerization and other post-translational modifications could favor the transition of these proteins into pathological conformations triggering their aggregation.…”

Section: Discussionmentioning

confidence: 99%

Zinc binding to RNA recognition motif of TDP-43 induces the formation of amyloid-like aggregates

Garnier

Devred

Byrne

et al. 2017

Sci Rep

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Aggregation of TDP-43 (transactive response DNA binding protein 43 kDa) is a hallmark of certain forms of amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD). Moreover, intracellular TDP-43-positive inclusions are often found in other neurodegenerative diseases. Recently it was shown that zinc ions can provoke the aggregation of endogenous TDP-43 in cells, allowing to assume a direct interaction of TDP-43 with zinc ions. In this work, we investigated zinc binding to the 102–269 TDP-43 fragment, which comprise the two RNA recognition motifs. Using isothermal titration calorimetry, mass spectrometry, and differential scanning fluorimetry, we showed that zinc binds to this TDP-43 domain with a dissociation constant in the micromolar range and modifies its tertiary structure leading to a decrease of its thermostability. Moreover, the study by dynamic light scattering and negative stain electron microscopy demonstrated that zinc ions induce auto-association process of this TDP-43 fragment into rope-like structures. These structures are thioflavin-T-positive allowing to hypothesize the direct implication of zinc ions in pathological aggregation of TDP-43.

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“…The protein concentration and timescales required for many types of measurements have resulted in tau-induced polymerization of tubulin even in the absence of additional assembly promoters (53). FCS allows for low tau concentrations (20 nM in our measurements), enabling the quantification of tubulin heterodimer binding under conditions where tubulin assembly is disfavored.…”

Section: Discussionmentioning

confidence: 99%

Tau mutants bind tubulin heterodimers with enhanced affinity

Elbaum‐Garfinkle

Cobb

Compton

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

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Tau is a microtubule binding protein that forms pathological aggregates in the brain in Alzheimer's disease and other tauopathies. Disease etiology is thought to arise from loss of native interactions between tau and microtubules, as well as from gain of toxicity tied to tau aggregation, although neither mechanism is well understood. Here we investigate the link between function and disease using disease-associated and disease-motivated mutants of tau. We find that mutations to highly conserved proline residues in repeats 2 and 3 of the microtubule binding domain have differential effects on tau binding to tubulin and the capacity of tau to enhance tubulin polymerization. Notably, mutations to these residues result in an increased affinity for tubulin dimers while having a negligible effect on binding to stabilized microtubules. We measure conformational changes in tau on binding to tubulin that provide a structural framework for the observed altered affinity and function. Additionally, we find that these mutations do not necessarily enhance aggregation, which could have important implications for tau therapeutic strategies that focus solely on searching for tau aggregation inhibitors. We propose a model that describes tau binding to tubulin dimers and a mechanism by which disease-relevant alterations to tau impact its function. Together, these results draw attention to the interaction between tau and free tubulin as playing an important role in mechanisms of tau pathology.

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New insights into tau–microtubules interaction revealed by isothermal titration calorimetry

Cited by 25 publications

References 19 publications

Molecular Mechanisms of Tau Binding to Microtubule and its Role in Microtubule Dynamics in Live Cells

Molecular Mechanisms of Tau Binding to Microtubule and its Role in Microtubule Dynamics in Live Cells

Zinc binding to RNA recognition motif of TDP-43 induces the formation of amyloid-like aggregates

Tau mutants bind tubulin heterodimers with enhanced affinity

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