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Chromatin-Independent Nuclear Envelope Assembly Induced by Ran GTPase in Xenopus Egg Extracts
The nuclear envelope (NE) forms a controlled boundary between the cytoplasm and the nucleus of eukaryotic cells. To facilitate investigation of mechanisms controlling NE assembly, we developed a cell-free system made from Xenopus laevis eggs to study the process in the absence of chromatin. NEs inco...
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Published in: | Science (American Association for the Advancement of Science) 2000-05, Vol.288 (5470), p.1429-1432 |
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creator | Zhang, Chuanmao Clarke, Paul R. |
description | The nuclear envelope (NE) forms a controlled boundary between the cytoplasm and the nucleus of eukaryotic cells. To facilitate investigation of mechanisms controlling NE assembly, we developed a cell-free system made from Xenopus laevis eggs to study the process in the absence of chromatin. NEs incorporating nuclear pores were assembled around beads coated with the guanosine triphosphatase Ran, forming pseudo-nuclei that actively imported nuclear proteins. NE assembly required the cycling of guanine nucleotides on Ran and was promoted by RCC1, a nucleotide exchange factor recruited to beads by Ran-guanosine diphosphate (Ran-GDP). Thus, concentration of Ran-GDP followed by generation of Ran-GTP is sufficient to induce NE assembly. |
doi_str_mv | 10.1126/science.288.5470.1429 |
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To facilitate investigation of mechanisms controlling NE assembly, we developed a cell-free system made from Xenopus laevis eggs to study the process in the absence of chromatin. NEs incorporating nuclear pores were assembled around beads coated with the guanosine triphosphatase Ran, forming pseudo-nuclei that actively imported nuclear proteins. NE assembly required the cycling of guanine nucleotides on Ran and was promoted by RCC1, a nucleotide exchange factor recruited to beads by Ran-guanosine diphosphate (Ran-GDP). Thus, concentration of Ran-GDP followed by generation of Ran-GTP is sufficient to induce NE assembly.</description><identifier>ISSN: 0036-8075</identifier><identifier>EISSN: 1095-9203</identifier><identifier>DOI: 10.1126/science.288.5470.1429</identifier><identifier>PMID: 10827954</identifier><identifier>CODEN: SCIEAS</identifier><language>eng</language><publisher>Washington, DC: American Society for the Advancement of Science</publisher><subject>Animals ; Biological and medical sciences ; Biological Transport, Active ; Cell Cycle Proteins ; Cell Extracts ; Cell nucleus ; Cell structures and functions ; Cells ; Cellular biology ; Chromatin ; Chromatin - metabolism ; Disease outbreaks ; DNA-Binding Proteins - metabolism ; Eggs ; Fluorescent antibody techniques ; Fundamental and applied biological sciences. Psychology ; Guanine Nucleotide Exchange Factors ; Guanosine Diphosphate - metabolism ; Guanosine Triphosphate - metabolism ; Imports ; Intermediate Filament Proteins ; Lamin Type B ; Membrane Fusion ; Molecular and cellular biology ; Molecular biology ; Nipah virus ; Nuclear Envelope - metabolism ; Nuclear Envelope - ultrastructure ; nuclear envelopes ; Nuclear membrane ; Nuclear membranes ; Nuclear pore complex proteins ; Nuclear Proteins - metabolism ; Nucleoplasmins ; Ova ; Ovum ; Phosphoproteins - metabolism ; Proteins ; ran GTP-Binding Protein - metabolism ; Ran protein ; Recombinant Fusion Proteins - metabolism ; Swine ; Telophase ; Viruses ; Xenopus ; Xenopus laevis ; Xenopus Proteins</subject><ispartof>Science (American Association for the Advancement of Science), 2000-05, Vol.288 (5470), p.1429-1432</ispartof><rights>Copyright 2000 American Association for the Advancement of Science</rights><rights>2000 INIST-CNRS</rights><rights>COPYRIGHT 2000 American Association for the Advancement of Science</rights><rights>COPYRIGHT 2000 American Association for the Advancement of Science</rights><rights>Copyright American Association for the Advancement of Science May 26, 2000</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c751t-859e77f0e7323e52c0533ab1e867daf7386a4a22b2dd9fff1fbb6f34d152885b3</citedby><cites>FETCH-LOGICAL-c751t-859e77f0e7323e52c0533ab1e867daf7386a4a22b2dd9fff1fbb6f34d152885b3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.proquest.com/docview/213596642/fulltextPDF?pq-origsite=primo$$EPDF$$P50$$Gproquest$$H</linktopdf><linktohtml>$$Uhttps://www.proquest.com/docview/213596642?pq-origsite=primo$$EHTML$$P50$$Gproquest$$H</linktohtml><link.rule.ids>314,777,781,2871,2872,21359,21375,27905,27906,33592,33593,33858,33859,43714,43861,58219,58452,73970,74146</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=1491606$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/10827954$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Zhang, Chuanmao</creatorcontrib><creatorcontrib>Clarke, Paul R.</creatorcontrib><title>Chromatin-Independent Nuclear Envelope Assembly Induced by Ran GTPase in Xenopus Egg Extracts</title><title>Science (American Association for the Advancement of Science)</title><addtitle>Science</addtitle><description>The nuclear envelope (NE) forms a controlled boundary between the cytoplasm and the nucleus of eukaryotic cells. 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To facilitate investigation of mechanisms controlling NE assembly, we developed a cell-free system made from Xenopus laevis eggs to study the process in the absence of chromatin. NEs incorporating nuclear pores were assembled around beads coated with the guanosine triphosphatase Ran, forming pseudo-nuclei that actively imported nuclear proteins. NE assembly required the cycling of guanine nucleotides on Ran and was promoted by RCC1, a nucleotide exchange factor recruited to beads by Ran-guanosine diphosphate (Ran-GDP). Thus, concentration of Ran-GDP followed by generation of Ran-GTP is sufficient to induce NE assembly.</abstract><cop>Washington, DC</cop><pub>American Society for the Advancement of Science</pub><pmid>10827954</pmid><doi>10.1126/science.288.5470.1429</doi><tpages>4</tpages></addata></record> |
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subjects | Animals Biological and medical sciences Biological Transport, Active Cell Cycle Proteins Cell Extracts Cell nucleus Cell structures and functions Cells Cellular biology Chromatin Chromatin - metabolism Disease outbreaks DNA-Binding Proteins - metabolism Eggs Fluorescent antibody techniques Fundamental and applied biological sciences. Psychology Guanine Nucleotide Exchange Factors Guanosine Diphosphate - metabolism Guanosine Triphosphate - metabolism Imports Intermediate Filament Proteins Lamin Type B Membrane Fusion Molecular and cellular biology Molecular biology Nipah virus Nuclear Envelope - metabolism Nuclear Envelope - ultrastructure nuclear envelopes Nuclear membrane Nuclear membranes Nuclear pore complex proteins Nuclear Proteins - metabolism Nucleoplasmins Ova Ovum Phosphoproteins - metabolism Proteins ran GTP-Binding Protein - metabolism Ran protein Recombinant Fusion Proteins - metabolism Swine Telophase Viruses Xenopus Xenopus laevis Xenopus Proteins |
title | Chromatin-Independent Nuclear Envelope Assembly Induced by Ran GTPase in Xenopus Egg Extracts |
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