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dc.contributor.authorZhang, Chuanmao
dc.contributor.authorGoldberg, Martin W
dc.contributor.authorMoore, William J
dc.contributor.authorAllen, Terence D
dc.contributor.authorClarke, Paul R
dc.date.accessioned2009-10-16T10:50:50Z
dc.date.available2009-10-16T10:50:50Z
dc.date.issued2002-11
dc.identifier.citationConcentration of Ran on chromatin induces decondensation, nuclear envelope formation and nuclear pore complex assembly. 2002, 81 (11):623-33 Eur. J. Cell Biol.en
dc.identifier.issn0171-9335
dc.identifier.pmid12494999
dc.identifier.urihttp://hdl.handle.net/10541/84333
dc.description.abstractNuclear envelope (NE) formation can be studied in a cell-free system made from Xenopus eggs. In this system, NE formation involves the small GTPase Ran. Ran associates with chromatin early in nuclear assembly and concentration of Ran on inert beads is sufficient to induce NE formation. Here, we show that Ran binds to chromatin prior to NE formation and recruits RCC1, the nucleotide exchange factor that generates Ran-GTP. In extracts prepared by high-speed centrifugation, increased concentrations of Ran are sufficient to induce chromatin decondensation and NE assembly. Using field emission in-lens scanning electron microscopy (FEISEM), we show that Ran promotes the formation of smoothed membranes and the assembly of nuclear pore complexes (NPCs). In contrast, RanT24N, a mutant that fails to bind GTP and inhibits RCC1, does not support efficient NE assembly, whereas RanQ69L, a mutant locked in a GTP-bound state, permits some membrane vesicle recruitment to chromatin, but inhibits vesicle fusion and NPC assembly. Thus, binding of Ran to chromatin, followed by local generation of Ran-GTP and GTP hydrolysis by Ran, induces chromatin decondensation, membrane vesicle recruitment, membrane formation and NPC assembly. We propose that the biological activity of Ran is determined by its targeting to structures such as chromatin as well as its guanine nucleotide bound state.
dc.language.isoenen
dc.subject.meshAnimals
dc.subject.meshBlotting, Western
dc.subject.meshCell Cycle Proteins
dc.subject.meshChromatin
dc.subject.meshFluorescent Antibody Technique
dc.subject.meshGuanine Nucleotide Exchange Factors
dc.subject.meshHumans
dc.subject.meshMicroscopy, Electron, Scanning
dc.subject.meshMutation
dc.subject.meshNuclear Envelope
dc.subject.meshNuclear Pore
dc.subject.meshNuclear Proteins
dc.subject.meshXenopus
dc.subject.meshXenopus Proteins
dc.subject.meshran GTP-Binding Protein
dc.titleConcentration of Ran on chromatin induces decondensation, nuclear envelope formation and nuclear pore complex assembly.en
dc.typeArticleen
dc.contributor.departmentDepartment of Structural Cell Biology, Paterson Institute for Cancer Research, Christie Hopsital, Masnchester M20 4BX, UKen
dc.identifier.journalEuropean Journal of Cell Biologyen
html.description.abstractNuclear envelope (NE) formation can be studied in a cell-free system made from Xenopus eggs. In this system, NE formation involves the small GTPase Ran. Ran associates with chromatin early in nuclear assembly and concentration of Ran on inert beads is sufficient to induce NE formation. Here, we show that Ran binds to chromatin prior to NE formation and recruits RCC1, the nucleotide exchange factor that generates Ran-GTP. In extracts prepared by high-speed centrifugation, increased concentrations of Ran are sufficient to induce chromatin decondensation and NE assembly. Using field emission in-lens scanning electron microscopy (FEISEM), we show that Ran promotes the formation of smoothed membranes and the assembly of nuclear pore complexes (NPCs). In contrast, RanT24N, a mutant that fails to bind GTP and inhibits RCC1, does not support efficient NE assembly, whereas RanQ69L, a mutant locked in a GTP-bound state, permits some membrane vesicle recruitment to chromatin, but inhibits vesicle fusion and NPC assembly. Thus, binding of Ran to chromatin, followed by local generation of Ran-GTP and GTP hydrolysis by Ran, induces chromatin decondensation, membrane vesicle recruitment, membrane formation and NPC assembly. We propose that the biological activity of Ran is determined by its targeting to structures such as chromatin as well as its guanine nucleotide bound state.


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