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A Rae1-containing ribonucleoprotein complex is required for mitotic spindle assembly.

Michael D Blower·Maxence Nachury·Rebecca Heald·Karsten Weis
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Abstract

Centrosome-independent microtubule polymerization around chromosomes has been shown to require a local gradient of RanGTP, which discharges mitotic cargoes from the nuclear import receptor importin beta. Here, we have used an activity-based assay in Xenopus egg extracts to purify the mRNA export protein Rae1 as a spindle assembly factor regulated by this pathway. Rae1 is a microtubule-associated protein that binds directly to importin beta. Depletion of Rae1 from extracts or cells severely inhibits mitotic spindle assembly. A purified Rae1 complex stabilizes microtubules in egg extracts in a RanGTP/importin beta-regulated manner. Interestingly, Rae1 exists in a large ribonucleoprotein complex, which requires RNA for its activity to control microtubule dynamics in vitro. Furthermore, we provide evidence that RNA associates with the mitotic spindle and that it plays a direct, translation-independent role in spindle assembly. Our studies reveal an unexpected function for RNA in spindle morphogenesis.

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AnimalsCell ExtractsFemaleHeLa CellsHumansMaleMicrotubulesNuclear Matrix-Associated ProteinsNuclear Pore Complex ProteinsNucleocytoplasmic Transport ProteinsOocytesRNA, MessengerRibonucleoproteinsSpermatozoaSpindle ApparatusXenopus ProteinsXenopus laevisbeta Karyopherinsran GTP-Binding ProteinJournal ArticleResearch Support, Non-U.S. Gov'tResearch Support, U.S. Gov't, P.H.S.

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Michael D Blower, Maxence Nachury, Rebecca Heald, Karsten Weis. 2005-04-22. A Rae1-containing ribonucleoprotein complex is required for mitotic spindle assembly.. https://doi.org/10.1016/j.cell.2005.02.016

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