We recently developed an assay in which nuclear export of the shuttling transcription factor NFAT (nuclear factor of activated T cells) can be reconstituted in permeabilized cells with the GTPase Ran and the nuclear export receptor CRM1. We have now used this assay to identify another export factor. After preincubation of permeabilized cells with a Ran mutant that cannot hydrolyze GTP (RanQ69L), cytosol supports NFAT export, but CRM1 and Ran alone do not. The RanQ69L preincubation leads to accumulation of CRM1 at the cytoplasmic periphery of the nuclear pore complex (NPC) in association with the p62 complex and Can/Nup214. RanGTP-dependent association of CRM1 with these nucleoporins was reconstituted in vitro. By biochemical fractionation and reconstitution, we showed that RanBP1 restores nuclear export after the RanQ69L preincubation. It also stimulates nuclear export in cells that have not been preincubated with RanQ69L. RanBP1 as well as Ran-binding domains of the cytoplasmic nucleoporin RanBP2 promote the release of CRM1 from the NPC. Taken together, our results indicate that RanGTP is important for the targeting of export complexes to the cytoplasmic side of the NPC and that RanBP1 and probably RanBP2 are involved in the dissociation of nuclear export complexes from the NPC in a terminal step of transport.
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17 May 1999
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May 17 1999
A Role for RanBP1 in the Release of CRM1 from the Nuclear Pore Complex in a Terminal Step of Nuclear Export
Ralph H. Kehlenbach,
Ralph H. Kehlenbach
Departments of Cell Biology and Molecular Biology, The Scripps Research Institute, La Jolla, California 92037
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Achim Dickmanns,
Achim Dickmanns
Departments of Cell Biology and Molecular Biology, The Scripps Research Institute, La Jolla, California 92037
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Angelika Kehlenbach,
Angelika Kehlenbach
Departments of Cell Biology and Molecular Biology, The Scripps Research Institute, La Jolla, California 92037
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Tinglu Guan,
Tinglu Guan
Departments of Cell Biology and Molecular Biology, The Scripps Research Institute, La Jolla, California 92037
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Larry Gerace
Larry Gerace
Departments of Cell Biology and Molecular Biology, The Scripps Research Institute, La Jolla, California 92037
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Ralph H. Kehlenbach
Departments of Cell Biology and Molecular Biology, The Scripps Research Institute, La Jolla, California 92037
Achim Dickmanns
Departments of Cell Biology and Molecular Biology, The Scripps Research Institute, La Jolla, California 92037
Angelika Kehlenbach
Departments of Cell Biology and Molecular Biology, The Scripps Research Institute, La Jolla, California 92037
Tinglu Guan
Departments of Cell Biology and Molecular Biology, The Scripps Research Institute, La Jolla, California 92037
Larry Gerace
Departments of Cell Biology and Molecular Biology, The Scripps Research Institute, La Jolla, California 92037
Address correspondence to Larry Gerace, Departments of Cell and Molecular Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037. Tel.: (619) 784-8514. Fax: (619) 784-9132. E-mail: [email protected]
Dr. Dickmanns' present address is Max Planck Institut für Biochemie, Am Klopferspitz 18a, D-82152 Martinsried, Germany.
Received:
December 30 1998
Revision Received:
March 11 1999
Online ISSN: 1540-8140
Print ISSN: 0021-9525
1999
J Cell Biol (1999) 145 (4): 645–657.
Article history
Received:
December 30 1998
Revision Received:
March 11 1999
Citation
Ralph H. Kehlenbach, Achim Dickmanns, Angelika Kehlenbach, Tinglu Guan, Larry Gerace; A Role for RanBP1 in the Release of CRM1 from the Nuclear Pore Complex in a Terminal Step of Nuclear Export . J Cell Biol 17 May 1999; 145 (4): 645–657. doi: https://doi.org/10.1083/jcb.145.4.645
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