Journal
JOURNAL OF CELL SCIENCE
Volume 126, Issue 13, Pages 2810-2819Publisher
COMPANY OF BIOLOGISTS LTD
DOI: 10.1242/jcs.120832
Keywords
Microtubule-associated protein Tau; Tubulin; Interaction; Fluorescence; Cell
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Funding
- INCa (Institut National du Cancer)
- Inserm (Institut National de la Sante et de la Recherche Medicale)
- AMU (Aix-Marseille Universite)
- DGOS [SIRIC label (Site de Recherche Integre sur le Cancer)]
- Region Provence Alps-Cote d'Azur
- ONET Technologies
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Despite 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 Forster 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 alpha-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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