2026-08-29T10:37:55Zhttps://repisalud.isciii.es/rest/oai/request

oai:repisalud.isciii.es:20.500.12105/230862024-11-29T17:01:10Zcom_20.500.12105_2173com_20.500.12105_2051col_20.500.12105_19597

00925njm 22002777a 4500 dc Zimmermann, Fabian author Serna, Marina author Ezquerra, Artur author Fernandez-Leiro, Rafael author Llorca Blanco, Oscar Antonio author Luders, Jens author Zimmermann, Fabian author Ezquerra, Artur author Fernandez-Leiro, Rafael author Luders, Jens author 2020-12-18 The microtubule nucleator ?-tubulin ring complex (?TuRC) is essential for the function of microtubule organizing centers such as the centrosome. Since its discovery over two decades ago, ?TuRC has evaded in vitro reconstitution and thus detailed structure-function studies. Here, we show that a complex of RuvB-like protein 1 (RUVBL1) and RUVBL2 "RUVBL" controls assembly and composition of ?TuRC in human cells. Likewise, RUVBL assembles ?TuRC from a minimal set of core subunits in a heterologous coexpression system. RUVBL interacts with ?TuRC subcomplexes but is not part of fully assembled ?TuRC. Purified, reconstituted ?TuRC has nucleation activity and resembles native ?TuRC as revealed by its cryo-electron microscopy (cryo-EM) structure at ~4.0-� resolution. We further use cryo-EM to identify features that determine the intricate, higher-order ?TuRC architecture. Our work finds RUVBL as an assembly factor that regulates ?TuRC in cells and allows production of recombinant ?TuRC for future in-depth mechanistic studies. Sci Adv . 2020 ;6(51):eabe0894. 10.1126/sciadv.abe0894 2375-2548 Science advances 33355144 https://hdl.handle.net/20.500.12105/23086 Assembly of the asymmetric human ?-tubulin ring complex by RUVBL1-RUVBL2 AAA ATPase.