The Distance between N and C Termini of Tau and of FTDP-17 Mutants Is Modulated by Microtubule Interactions in Living Cells
The microtubule (MT)-associated protein Tau is a natively unfolded protein, involved in a number of neurodegenerative disorders, collectively called tauopathies, aggregating in neurofibrillary tangles (NFT). It is an open question how the conversion from a MT bound molecule to an aggregation-prone T...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2017-06-01
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| Series: | Frontiers in Molecular Neuroscience |
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| Online Access: | http://journal.frontiersin.org/article/10.3389/fnmol.2017.00210/full |
| _version_ | 1818061882308689920 |
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| author | Cristina Di Primio Valentina Quercioli Giacomo Siano Matteo Rovere Branislav Kovacech Michal Novak Antonino Cattaneo |
| author_facet | Cristina Di Primio Valentina Quercioli Giacomo Siano Matteo Rovere Branislav Kovacech Michal Novak Antonino Cattaneo |
| author_sort | Cristina Di Primio |
| collection | DOAJ |
| description | The microtubule (MT)-associated protein Tau is a natively unfolded protein, involved in a number of neurodegenerative disorders, collectively called tauopathies, aggregating in neurofibrillary tangles (NFT). It is an open question how the conversion from a MT bound molecule to an aggregation-prone Tau species occurs and, also, if and how tauopathy-related mutations affect its behavior in the cell. To address these points, we exploited a genetically encoded FRET sensor based on the full length Tau protein, to monitor in real time Tau conformational changes in different conditions in live cells. By studying the FRET signal we found that soluble Tau molecules, detached from MTs, display an unfolded structure. On the contrary, we observed an increased FRET signal generated by Tau monomers bound to MT, indicating that the association with MTs induced a folding of Tau protein, decreasing the distance between its N and C termini. We exploited the FRET sensor to investigate the impact of FTDP-17 mutations and of phosphorylation-site mutations on Tau folding and mobility in live cells. We demonstrated that the FTDP-17 Tau mutations weaken the interaction of Tau with cellular MTs, shifting the equilibrium towards the soluble pool while, conversely, phosphorylation site mutations shift the equilibrium of Tau towards the MT-bound state and a more closed conformation. |
| first_indexed | 2024-12-10T13:55:22Z |
| format | Article |
| id | doaj.art-91b173261d714619aa224d56b03e4cc9 |
| institution | Directory Open Access Journal |
| issn | 1662-5099 |
| language | English |
| last_indexed | 2024-12-10T13:55:22Z |
| publishDate | 2017-06-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Molecular Neuroscience |
| spelling | doaj.art-91b173261d714619aa224d56b03e4cc92022-12-22T01:46:01ZengFrontiers Media S.A.Frontiers in Molecular Neuroscience1662-50992017-06-011010.3389/fnmol.2017.00210272141The Distance between N and C Termini of Tau and of FTDP-17 Mutants Is Modulated by Microtubule Interactions in Living CellsCristina Di Primio0Valentina Quercioli1Giacomo Siano2Matteo Rovere3Branislav Kovacech4Michal Novak5Antonino Cattaneo6Bio@SNS Laboratory, Scuola Normale SuperiorePisa, ItalyBio@SNS Laboratory, Scuola Normale SuperiorePisa, ItalyBio@SNS Laboratory, Scuola Normale SuperiorePisa, ItalyBio@SNS Laboratory, Scuola Normale SuperiorePisa, ItalyInstitute of Neuroimmunology, Slovak Academy of Sciences, Axon Neuroscience SEBratislava, SlovakiaInstitute of Neuroimmunology, Slovak Academy of Sciences, Axon Neuroscience SEBratislava, SlovakiaBio@SNS Laboratory, Scuola Normale SuperiorePisa, ItalyThe microtubule (MT)-associated protein Tau is a natively unfolded protein, involved in a number of neurodegenerative disorders, collectively called tauopathies, aggregating in neurofibrillary tangles (NFT). It is an open question how the conversion from a MT bound molecule to an aggregation-prone Tau species occurs and, also, if and how tauopathy-related mutations affect its behavior in the cell. To address these points, we exploited a genetically encoded FRET sensor based on the full length Tau protein, to monitor in real time Tau conformational changes in different conditions in live cells. By studying the FRET signal we found that soluble Tau molecules, detached from MTs, display an unfolded structure. On the contrary, we observed an increased FRET signal generated by Tau monomers bound to MT, indicating that the association with MTs induced a folding of Tau protein, decreasing the distance between its N and C termini. We exploited the FRET sensor to investigate the impact of FTDP-17 mutations and of phosphorylation-site mutations on Tau folding and mobility in live cells. We demonstrated that the FTDP-17 Tau mutations weaken the interaction of Tau with cellular MTs, shifting the equilibrium towards the soluble pool while, conversely, phosphorylation site mutations shift the equilibrium of Tau towards the MT-bound state and a more closed conformation.http://journal.frontiersin.org/article/10.3389/fnmol.2017.00210/fullTaubiosensorconformationFRETFRAPmutation |
| spellingShingle | Cristina Di Primio Valentina Quercioli Giacomo Siano Matteo Rovere Branislav Kovacech Michal Novak Antonino Cattaneo The Distance between N and C Termini of Tau and of FTDP-17 Mutants Is Modulated by Microtubule Interactions in Living Cells Frontiers in Molecular Neuroscience Tau biosensor conformation FRET FRAP mutation |
| title | The Distance between N and C Termini of Tau and of FTDP-17 Mutants Is Modulated by Microtubule Interactions in Living Cells |
| title_full | The Distance between N and C Termini of Tau and of FTDP-17 Mutants Is Modulated by Microtubule Interactions in Living Cells |
| title_fullStr | The Distance between N and C Termini of Tau and of FTDP-17 Mutants Is Modulated by Microtubule Interactions in Living Cells |
| title_full_unstemmed | The Distance between N and C Termini of Tau and of FTDP-17 Mutants Is Modulated by Microtubule Interactions in Living Cells |
| title_short | The Distance between N and C Termini of Tau and of FTDP-17 Mutants Is Modulated by Microtubule Interactions in Living Cells |
| title_sort | distance between n and c termini of tau and of ftdp 17 mutants is modulated by microtubule interactions in living cells |
| topic | Tau biosensor conformation FRET FRAP mutation |
| url | http://journal.frontiersin.org/article/10.3389/fnmol.2017.00210/full |
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