GTP hydrolysis is required for vesicle fusion during nuclear envelope assembly in vitro.

作者: A L Boman , M R Delannoy , K L Wilson

DOI: 10.1083/JCB.116.2.281

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摘要: Nuclear envelope assembly was studied in vitro using extracts from Xenopus eggs. Nuclear-specific vesicles bound to demembranated sperm chromatin but did not fuse the absence of cytosol. Addition cytosol stimulated vesicle fusion, pore complex assembly, and eventual nuclear growth. Vesicle binding fusion were assayed by light electron microscopy. ATP GTP caused limited enclosure observed. This result suggested that nondialyzable soluble components required for fusion. gamma S guanylyl imidodiphosphate significantly inhibited had no effect on chromatin. Preincubation membranes with 1 mM or impair when fresh However, preincubation plus irreversible inhibition The factor mediating S, which we named GTP-dependent (GSF), titratable depleted incubation excess suggesting a stoichiometric interaction between GSF membrane component presence S. In preliminary experiments, remained active chromatin-bound vesicles, may be reaction itself. We propose hydrolysis is at step before vesicles.

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