Biochemical reconstitutions reveal principles of human γ-TuRC assembly and function
| العنوان: | Biochemical reconstitutions reveal principles of human γ-TuRC assembly and function |
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| المؤلفون: | Kelly R. Molloy, Tarun M. Kapoor, Michal Wieczorek, Amol Aher, Linas Urnavicius, Shih-Chieh Ti, Brian T. Chait |
| المصدر: | The Journal of Cell Biology The Journal of Cell Biology, 220 (3) |
| بيانات النشر: | ETH Zurich, 2021. |
| سنة النشر: | 2021 |
| مصطلحات موضوعية: | 0303 health sciences, Guanine, Extramural, Kinetics, Cell Biology, Biology, Biochemistry, Microtubules, In vitro, 03 medical and health sciences, chemistry.chemical_compound, 0302 clinical medicine, Tubulin, chemistry, Structural Biology, Microtubule, Report, biology.protein, Biophysics, Cytoskeleton, Microtubule-Organizing Center, 030217 neurology & neurosurgery, Function (biology), Actin, 030304 developmental biology |
| الوصف: | The formation of cellular microtubule networks is regulated by the γ-tubulin ring complex (γ-TuRC). This ∼2.3 MD assembly of >31 proteins includes γ-tubulin and GCP2-6, as well as MZT1 and an actin-like protein in a “lumenal bridge” (LB). The challenge of reconstituting the γ-TuRC has limited dissections of its assembly and function. Here, we report a biochemical reconstitution of the human γ-TuRC (γ-TuRC-GFP) as a ∼35 S complex that nucleates microtubules in vitro. In addition, we generate a subcomplex, γ-TuRCΔLB-GFP, which lacks MZT1 and actin. We show that γ-TuRCΔLB-GFP nucleates microtubules in a guanine nucleotide–dependent manner and with similar efficiency as the holocomplex. Electron microscopy reveals that γ-TuRC-GFP resembles the native γ-TuRC architecture, while γ-TuRCΔLB-GFP adopts a partial cone shape presenting only 8–10 γ-tubulin subunits and lacks a well-ordered lumenal bridge. Our results show that the γ-TuRC can be reconstituted using a limited set of proteins and suggest that the LB facilitates the self-assembly of regulatory interfaces around a microtubulenucleating “core” in the holocomplex. The Journal of Cell Biology, 220 (3) ISSN:0021-9525 ISSN:1540-8140 |
| وصف الملف: | application/application/pdf |
| اللغة: | English |
| تدمد: | 0021-9525 1540-8140 |
| DOI: | 10.3929/ethz-b-000515975 |
| الوصول الحر: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::a51c28d65428384ffcdd705dd74b52d5Test |
| حقوق: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....a51c28d65428384ffcdd705dd74b52d5 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 00219525 15408140 |
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| DOI: | 10.3929/ethz-b-000515975 |