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PMID: 8896452 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Identification of different roles for RanGDP and RanGTP in nuclear protein import.

The EMBO journal ·Vol. 15 ·No. 20 ·1996-10-15 ·Pages 5584-94

Görlich D, Panté N, Kutay U, Aebi U, Bischoff FR

Abstract

The importin-alpha/beta heterodimer and the GTPase Ran play key roles in nuclear protein import. Importin binds the nuclear localization signal (NLS). Translocation of the resulting import ligand complex through the nuclear pore complex (NPC) requires Ran and is terminated at the nucleoplasmic side by its disassembly. The principal GTP exchange factor for Ran is the nuclear protein RCC1, whereas the major RanGAP is cytoplasmic, predicting that nuclear Ran is mainly in the GTP form and cytoplasmic Ran is in the GDP-bound form. Here, we show that nuclear import depends on cytoplasmic RanGDP and free GTP, and that RanGDP binds to the NPC. Therefore, import might involve nucleotide exchange and GTP hydrolysis on NPC-bound Ran. RanGDP binding to the NPC is not mediated by the Ran binding sites of importin-beta, suggesting that translocation is not driven from these sites. Consistently, a mutant importin-beta deficient in Ran binding can deliver its cargo up to the nucleoplasmic side of the NPC. However, the mutant is unable to release the import substrate into the nucleoplasm. Thus, binding of nucleoplasmic RanGTP to importin-beta probably triggers termination, i.e. the dissociation of importin-alpha from importin-beta and the subsequent release of the import substrate into the nucleoplasm.

MeSH Terms
Animals Biological Transport Carrier Proteins/metabolism Cell Nucleus/metabolism GTP-Binding Proteins/metabolism GTPase-Activating Proteins Guanosine Diphosphate/metabolism Guanosine Triphosphate/metabolism HeLa Cells Humans Mice Microinjections Nuclear Proteins/metabolism Nucleosomes/metabolism Saccharomyces cerevisiae Proteins Xenopus Xenopus Proteins alpha Karyopherins beta Karyopherins ran GTP-Binding Protein
Chemicals
Carrier Proteins GTPase-Activating Proteins Nuclear Proteins Nucleosomes RANGAP1 protein, human RNA1 protein, S cerevisiae RanGAP1 protein, Xenopus Saccharomyces cerevisiae Proteins Xenopus Proteins alpha Karyopherins beta Karyopherins Guanosine Diphosphate Guanosine Triphosphate GTP-Binding Proteins ran GTP-Binding Protein
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Görlich D
Zentrum für Molekulare Biologie der Universität Heidelberg, Germany.
Panté N
Kutay U
Aebi U
Bischoff F R
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1996-10-15
Pages
5584-94
Language
English
Region
England
NLM ID
8208664
PMCID
PMC452303
Subset
IM
External Links
PubMed source PMC Full Text View journal details
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