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RanGTP and importin β regulate meiosis I spindle assembly and function in mouse oocytes

D. Drutovic, X. Duan, R. Li, P. Kalab, P. Solc,

. 2020 ; 39 (1) : e101689. [pub] 20191016

Language English Country Great Britain

Document type Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't

Persistent link   https://www.medvik.cz/link/bmc20025273

Grant support
LO1609 National Sustainability Program of the Czech Ministry of Education, Youth and Sports (MEYS) - International
LTAUSA17097 National Sustainability Program of the Czech Ministry of Education, Youth and Sports (MEYS) - International
R01 HD086577 NICHD NIH HHS - United States
S10 OD020007 NIH HHS - United States

Homologous chromosome segregation during meiosis I (MI) in mammalian oocytes is carried out by the acentrosomal MI spindles. Whereas studies in human oocytes identified Ran GTPase as a crucial regulator of the MI spindle function, experiments in mouse oocytes questioned the generality of this notion. Here, we use live-cell imaging with fluorescent probes and Förster resonance energy transfer (FRET) biosensors to monitor the changes in Ran and importin β signaling induced by perturbations of Ran in mouse oocytes while examining the MI spindle dynamics. We show that unlike RanT24N employed in previous studies, a RanT24N, T42A double mutant inhibits RanGEF without perturbing cargo binding to importin β and disrupts MI spindle function in chromosome segregation. Roles of Ran and importin β in the coalescence of microtubule organizing centers (MTOCs) and MI spindle assembly are further supported by the use of the chemical inhibitor importazole, whose effects are partially rescued by the GTP hydrolysis-resistant RanQ69L mutant. These results indicate that RanGTP is essential for MI spindle assembly and function both in humans and mice.

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