Concentration of Ran on chromatin induces decondensation, nuclear envelope formation and nuclear pore complex assembly.

2.50
Hdl Handle:
http://hdl.handle.net/10541/84333
Title:
Concentration of Ran on chromatin induces decondensation, nuclear envelope formation and nuclear pore complex assembly.
Authors:
Zhang, Chuanmao; Goldberg, Martin W; Moore, William J; Allen, Terence D; Clarke, Paul R
Abstract:
Nuclear 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.
Affiliation:
Department of Structural Cell Biology, Paterson Institute for Cancer Research, Christie Hopsital, Masnchester M20 4BX, UK
Citation:
Concentration of Ran on chromatin induces decondensation, nuclear envelope formation and nuclear pore complex assembly. 2002, 81 (11):623-33 Eur. J. Cell Biol.
Journal:
European Journal of Cell Biology
Issue Date:
Nov-2002
URI:
http://hdl.handle.net/10541/84333
PubMed ID:
12494999
Type:
Article
Language:
en
ISSN:
0171-9335
Appears in Collections:
All Paterson Institute for Cancer Research

Full metadata record

DC FieldValue Language
dc.contributor.authorZhang, Chuanmaoen
dc.contributor.authorGoldberg, Martin Wen
dc.contributor.authorMoore, William Jen
dc.contributor.authorAllen, Terence Den
dc.contributor.authorClarke, Paul Ren
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.en
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

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