Tau is an intrinsically disordered microtubule-associated protein (MAP) implicated in neurodegenerative disease. On microtubules, tau molecules segregate into two kinetically distinct phases, consisting of either independently diffusing molecules or interacting molecules that form cohesive ‘envelopes’ around microtubules. Envelopes differentially regulate lattice accessibility for other MAPs, but the mechanism of envelope formation remains unclear. Here we find that tau envelopes form cooperatively, locally altering the spacing of tubulin dimers within the microtubule lattice. Envelope formation compacted the underlying lattice, whereas lattice extension induced tau envelope disassembly. Investigating other members of the tau family, we find that MAP2 similarly forms envelopes governed by lattice spacing, whereas MAP4 cannot. Envelopes differentially biased motor protein movement, suggesting that tau family members could spatially divide the microtubule surface into functionally distinct regions. We conclude that the interdependent allostery between lattice spacing and cooperative envelope formation provides the molecular basis for spatial regulation of microtubule-based processes by tau and MAP2.
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The authors thank members of the MOM lab for feedback and discussion during the project and the Protein Facility of MPI-CBG and V. Váňová for technical support. We also thank A. Carter for critical reading of the manuscript, O. Kučera for establishing the optical tweezers assay and J. Sabó for support in obtaining imaging data. This work was supported by Czech Science Foundation grant 19–27477X to Z.L. and L.L. and grant 20-04068S to M.B. We acknowledge the financial support from the Charles University Grant Agency (GAUK no. 373821 to V.S.) and the project ‘Grant Schemes at CU’ (reg. no. CZ.02.2.69/0.0/0.0/19_073/0016935) to T.H. and V.S., grants 1R35GM124889 to R.J.M. and 1R35GM133688 to K.M.O.M. K.M.O.M. is also supported by the Pew Charitable Trusts grant A19-0406. We acknowledge the institutional support from the CAS (RVO: 86652036), CMS supported by MEYS CR (LM2015043), and the Imaging Methods Core Facility at BIOCEV, an institution supported by the MEYS CR (Large RI Project LM2018129 Czech-BioImaging) and ERDF (project no. CZ.02.1.01/0.0/ 0.0/16_013/0001775) for their support in obtaining imaging data presented in this paper.
The authors declare no competing interests.
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Supplementary Figs 1–4, Supplementary Movie captions, Source data files, Supplementary Data files.
Video related to Fig. 1c. Structural changes in the microtubule lattice upon cooperative binding of tau. Time-lapse movie of tau-mCherry (green) envelopes growing on a taxol-lattice microtubule. Twenty nanomolar tau-mCherry is added to an initially bent microtubule. The microtubule straightens out when the tau envelope forms in the curved region. Scale bar, 2 µm.
Video related to Fig. 1e. Kinesin-1 walking on taxol- and GMPCPP-lattice microtubules in presence of elevated concentrations of tau. Time-lapse movie of kinesin-1 (magenta) stepping on taxol- and GMPCPP-lattice microtubules in the presence of elevated concentrations of tau (green). Sixty nanomolar kinesin-1-GFP was added to a taxol-lattice microtubule (top) and a GMPCPP-lattice microtubule (bottom) in the presence of 600 nM tau-mCherry. Processive movement of kinesin-1 was detected over the entire length of GMPCPP-lattice microtubules, while no processive movement of kinesin-1 was detected on taxol-lattice microtubules, indicating the presence of tau envelopes on taxol-lattice microtubules, and the absence of tau envelopes on GMPCPP-lattice microtubules even at these elevated concentrations of tau. Scale bar, 2 µm.
Video related to Fig. 1g. Compaction of a speckled-labeled microtubule upon cooperative binding of tau. Time-lapse movie of a speckled microtubule after the addition of tau. 200 nM tau-eGFP (green) was added to a speckled-labeled taxol-lattice microtubule (speckles in white). Upon cooperative binding of tau, speckles can be seen moving closer to each other, indicating the compaction of the microtubule lattice. Scale bar: 2 µm
Video related to Fig. 1i. SiR-tubulin dissociation of microtubule lattice within tau envelopes regions. Time-lapse movie of 2 μM SiR-tubulin (magenta) on the microtubule lattice after the addition of 20 nM tau-mCherry (green). At the positions where tau envelopes grow on the microtubule lattice (left), the density of SiR-tubulin locally decreases (middle). Right, overlay. Scale bar, 2 µm.
Video related to Fig. 2a. Local extension of GDP-lattice microtubule induces tau envelope disassembly in the presence of taxol. Time-lapse movie of 20 nM tau-mCherry (green) on a GMPCPP-capped GDP-lattice microtubule. At t = 0 s, tau is removed from the solution and microtubules are bent using hydrodynamic flow. During the hydrodynamic flow, imaging buffer with 10 μM taxol is added to the measurement chamber. In the presence of taxol, tau envelopes disassemble from their boundaries as well as from the location of the initially bent position. Scale bar, 5 µm.
Video related to Fig. 2a. Local extension of GDP-lattice microtubule does not induce tau envelope disassembly in the absence of taxol. Time-lapse movie of 20 nM tau-mCherry (green) on GMPCPP-capped GDP-lattice microtubules. At t = 0 s, tau is removed from the solution and microtubules are bent using hydrodynamic flow. During the hydrodynamic flow, imaging buffer without taxol is added to the measurement chamber. In the absence of taxol, no tau envelope disassembly was observed and disruptions in the tau envelopes owing to bending the microtubule lattice were found to reform within a 200 s timeframe. Scale bar, 5 µm.
Video related to Fig. 2g. Global extension of the microtubule lattice in the optical tweezers induces faster envelope disassembly. Time-lapse movie of the disassembly of tau envelopes on a microtubule suspended between two beads and stretched using external force. Initially, a single taxol-lattice microtubule is suspended between two beads and 60 nM tau-mCherry is added to the measurement chamber. The microtubule is then globally extended by slowly moving the beads apart until a set force of 40 pN is measured. Tau is then removed from solution so that envelope disassembly is observed. Global extension of the microtubule lattice in the optical tweezers induces faster envelope disassembly. Scale bar, 3 µm.
Video related to Fig. 2i. In the absence of global extension of the microtubule lattice in the optical tweezers, tau envelopes disassemble slower. Time-lapse movie of the disassembly of tau envelopes on a microtubule attached to a single bead and relaxed in the absence of external force. Initially, a single taxol-lattice microtubule is attached to a bead and 60 nM tau-mCherry is added to the measurement chamber. Tau is then removed from solution so that envelope disassembly is observed. In the absence of global extension of the microtubule lattice in the optical tweezers, tau envelopes disassemble slower. Scale bar, 3 µm.
Video related to Fig. 4a. Tau dissociates from microtubules in vivo upon addition of taxol. Time-lapse movie of a U-2 OS cell expressing eGFP-tau (white) after the addition of 0.01 μM taxol. Tau signal can be seen gradually dissociating from the microtubules in vivo after the addition of taxol. Scale bar, 10 µm.
Video related to Fig. 4a. Tau does not dissociate from microtubules after the addition of DMSO. Time-lapse movie of a U-2 OS cell expressing eGFP-tau (white) after the addition of DMSO in the absence of taxol. Tau remains on the microtubules in vivo in the control experiment in the absence of taxol. Scale bar, 10 µm.
Video related to Fig. 4c. MAP4 does not dissociate from microtubules in vivo upon addition of taxol. Time-lapse movie of a U-2 OS cell expressing eGFP-MAP4 after the addition of 0.01 μM taxol. In contrast to tau, MAP4 remains on the microtubules in vivo after the addition of taxol. Scale bar, 10 µm.
Video related to Fig. 4c. MAP4 does not dissociate from microtubules in vivo upon addition of DMSO. Time-lapse movie of a U-2 OS cell expressing eGFP-MAP4 after the addition of DMSO in the absence of taxol. MAP4 remains on the microtubules in vivo in the control experiment in the absence of taxol. Scale bar, 10 µm.
Video related to Fig. 4e. Tau dissociates from microtubules in vivo strongly resembling tau envelope disassembly in vitro. Time-lapse movie of a single microtubule in a U-2 OS cell expressing eGFP-tau after the addition of 0.01 μM taxol at t = 0 min. During the dissociation of tau from the microtubules, fissions appear in the tau density that disassemble from their boundaries, closely resembling tau envelope disassembly in vitro. Scale bar, 2 µm.
Video related to Supplementary Figure 4d. Tau dissociates from microtubules in vivo strongly resembling tau envelope disassembly in vitro. Time-lapse movie of a second example of a single microtubule in a U-2 OS cell expressing eGFP-tau after the addition of 0.01 μM taxol at t = 0 min. During the dissociation of tau from the microtubules, fissions appear in the tau density that disassemble from their boundaries, closely resembling tau envelope disassembly in vitro. Scale bar, 1 µm.
Video related to Supplementary Figure 4d. Tau dissociates from microtubules in vivo strongly resembling tau envelope disassembly in vitro. Time-lapse movie of a third example of a single microtubule in a U-2 OS cell expressing eGFP-tau after the addition of 0.01 μM taxol at t = 0 min. During the dissociation of tau from the microtubules, fissions appear in the tau density that disassemble from their boundaries, closely resembling tau envelope disassembly in vitro. Scale bar, 1 µm.
Statistical Source Data for Supplementary Figure 1.
Statistical Source Data for Supplementary Figure 2.
Statistical Source Data for Supplementary Figure 3.
Statistical Source Data for Supplementary Figure 4.
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Siahaan, V., Tan, R., Humhalova, T. et al. Microtubule lattice spacing governs cohesive envelope formation of tau family proteins. Nat Chem Biol 18, 1224–1235 (2022). https://doi.org/10.1038/s41589-022-01096-2
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