Chromatin-independent nuclear envelope assembly induced by Ran GTPase in Xenopus egg extracts

Chuanmao Zhang, Paul R. Clarke

Research output: Contribution to journalArticle

155 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)1429-1432
Number of pages4
JournalScience
Volume288
Issue number5470
DOIs
Publication statusPublished - 26 May 2000

Fingerprint

GTP Phosphohydrolases
Nuclear Envelope
Xenopus
Chromatin
Ovum
Guanosine
Diphosphates
Nuclear Pore
Guanine Nucleotides
Cell-Free System
Xenopus laevis
Eukaryotic Cells
Nuclear Proteins
Guanosine Triphosphate
Eggs
Cytoplasm
Nucleotides

Cite this

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abstract = "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.",
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Chromatin-independent nuclear envelope assembly induced by Ran GTPase in Xenopus egg extracts. / Zhang, Chuanmao; Clarke, Paul R.

In: Science, Vol. 288, No. 5470, 26.05.2000, p. 1429-1432.

Research output: Contribution to journalArticle

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