CLASP Mediates Microtubule Repair by Restricting Lattice Damage and Regulating Tubulin Incorporation
| Autor: | Aher, Amol, Rai, Dipti, Schaedel, Laura, Gaillard, Jeremie, John, Karin, Liu, Qingyang, Altelaar, Maarten, Blanchoin, Laurent, Thery, Manuel, Akhmanova, Anna, Celbiologie, Sub Cell Biology, Afd Biomol.Mass Spect. and Proteomics, Biomolecular Mass Spectrometry and Proteomics |
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| Přispěvatelé: | Celbiologie, Sub Cell Biology, Afd Biomol.Mass Spect. and Proteomics, Biomolecular Mass Spectrometry and Proteomics, Department of Biology, Faculty of Sciences, Utrecht University, CytoMorphoLab, Physiologie cellulaire et végétale (LPCV), Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Grenoble Alpes (UGA)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Grenoble Alpes (UGA), DYnamique des Fluides COmplexes et Morphogénèse [Grenoble] (DYFCOM-LIPhy ), Laboratoire Interdisciplinaire de Physique [Saint Martin d’Hères] (LIPhy ), Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes (UGA)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes (UGA), Bijvoet Center for Biomolecular Research [Utrecht], Utrecht University [Utrecht], Cell Biology Program, European Project: Synergy grant 609822, Université Grenoble Alpes (UGA)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes (UGA)-Centre National de la Recherche Scientifique (CNRS) |
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Microtubule disassembly in vitro reconstitution microtubule repair Motility microtubule rescue [SDV.BC]Life Sciences [q-bio]/Cellular Biology Biology Microtubules Article General Biochemistry Genetics and Molecular Biology 03 medical and health sciences 0302 clinical medicine Microtubule microtubule catastrophe Humans lattice defects Microtubule nucleation microtubule dynamics CLASP HEK293 Cells 030104 developmental biology Tubulin laser microsurgery tubulin Lattice defects TOG2 biology.protein Biophysics Laser microsurgery General Agricultural and Biological Sciences Microtubule-Associated Proteins 030217 neurology & neurosurgery Intracellular Protein Binding |
| Zdroj: | Current Biology, 30(11), 2175. Cell Press Current Biology-CB Current Biology-CB, 2020, 30 (11), pp.2175-2183. ⟨10.1016/j.cub.2020.03.070⟩ Current Biology Current Biology-CB, Elsevier, 2020, 30 (11), pp.2175-2183. ⟨10.1016/j.cub.2020.03.070⟩ |
| ISSN: | 0960-9822 1879-0445 |
| DOI: | 10.1016/j.cub.2020.03.070⟩ |
| Popis: | Summary Microtubules play a key role in cell division, motility, and intracellular trafficking. Microtubule lattices are generally regarded as stable structures that undergo turnover through dynamic instability of their ends [1]. However, recent evidence suggests that microtubules also exchange tubulin dimers at the sites of lattice defects, which can be induced by mechanical stress, severing enzymes, or occur spontaneously during polymerization [2, 3, 4, 5, 6]. Tubulin incorporation can restore microtubule integrity; moreover, “islands” of freshly incorporated GTP-tubulin can inhibit microtubule disassembly and promote rescues [3, 4, 6, 7, 8]. Microtubule repair occurs in vitro in the presence of tubulin alone [2, 3, 4, 5, 6, 9]. However, in cells, it is likely to be regulated by specific factors, the nature of which is currently unknown. CLASPs are interesting candidates for microtubule repair because they induce microtubule nucleation, stimulate rescue, and suppress catastrophes by stabilizing incomplete growing plus ends with lagging protofilaments and promoting their conversion into complete ones [10, 11, 12, 13, 14, 15, 16, 17]. Here, we used in vitro reconstitution assays combined with laser microsurgery and microfluidics to show that CLASP2α indeed stimulates microtubule lattice repair. CLASP2α promoted tubulin incorporation into damaged lattice sites, thereby restoring microtubule integrity. Furthermore, it induced the formation of complete tubes from partial protofilament assemblies and inhibited microtubule softening caused by hydrodynamic-flow-induced bending. The catastrophe-suppressing domain of CLASP2α, TOG2, combined with a microtubule-tethering region, was sufficient to stimulate microtubule repair, suggesting that catastrophe suppression and lattice repair are mechanistically similar. Our results suggest that the cellular machinery controlling microtubule nucleation and growth can also help to maintain microtubule integrity. Graphical Abstract Highlights • CLASP stabilizes damaged microtubule lattices • CLASP converts partial protofilament assemblies into complete tubes • CLASP promotes complete repair of microtubule lattice defects • CLASP inhibits softening of microtubules bent by hydrodynamic flow Using in vitro reconstitution assays combined with laser microsurgery and microfluidics, Aher et al. show that CLASP, a positive regulator of microtubule growth, promotes microtubule repair by stabilizing incomplete lattices and stimulating continuous tubulin incorporation into damaged sites. |
| Databáze: | OpenAIRE |
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