XB-ART-17568
EMBO J
1996 Oct 15;1520:5584-94.
Show Gene links
Show Anatomy links
Identification of different roles for RanGDP and RanGTP in nuclear protein import.
Görlich D
,
Panté N
,
Kutay U
,
Aebi U
,
Bischoff FR
.
???displayArticle.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.
???displayArticle.pubmedLink??? 8896452
???displayArticle.pmcLink??? PMC452303
Species referenced: Xenopus
Genes referenced: ran rcc1
References [+] :
Adam,
Nuclear protein import in permeabilized mammalian cells requires soluble cytoplasmic factors.
1990, Pubmed,
Xenbase
Adam, Nuclear protein import in permeabilized mammalian cells requires soluble cytoplasmic factors. 1990, Pubmed , Xenbase
Adam, Cytosolic proteins that specifically bind nuclear location signals are receptors for nuclear import. 1991, Pubmed
Adam, Identification of cytosolic factors required for nuclear location sequence-mediated binding to the nuclear envelope. 1994, Pubmed
Amberg, Nuclear PRP20 protein is required for mRNA export. 1993, Pubmed
Atkinson, Characterization of an essential Saccharomyces cerevisiae gene related to RNA processing: cloning of RNA1 and generation of a new allele with a novel phenotype. 1985, Pubmed
Baschong, Small colloidal gold conjugated to Fab fragments or to immunoglobulin G as high-resolution labels for electron microscopy: a technical overview. 1990, Pubmed
Becker, RNA1 encodes a GTPase-activating protein specific for Gsp1p, the Ran/TC4 homologue of Saccharomyces cerevisiae. 1995, Pubmed
Beddow, The Ran/TC4 GTPase-binding domain: identification by expression cloning and characterization of a conserved sequence motif. 1995, Pubmed
Bischoff, Catalysis of guanine nucleotide exchange on Ran by the mitotic regulator RCC1. 1991, Pubmed , Xenbase
Bischoff, Mitotic regulator protein RCC1 is complexed with a nuclear ras-related polypeptide. 1991, Pubmed
Bischoff, RanGAP1 induces GTPase activity of nuclear Ras-related Ran. 1994, Pubmed
Bischoff, Human RanGTPase-activating protein RanGAP1 is a homologue of yeast Rna1p involved in mRNA processing and transport. 1995, Pubmed
Bischoff, Co-activation of RanGTPase and inhibition of GTP dissociation by Ran-GTP binding protein RanBP1. 1995, Pubmed
Bischoff, Catalysis of guanine nucleotide exchange of Ran by RCC1 and stimulation of hydrolysis of Ran-bound GTP by Ran-GAP1. 1995, Pubmed
Carey, Evidence using a green fluorescent protein-glucocorticoid receptor chimera that the Ran/TC4 GTPase mediates an essential function independent of nuclear protein import. 1996, Pubmed
Cheng, Diverse effects of the guanine nucleotide exchange factor RCC1 on RNA transport. 1995, Pubmed
Chi, Sequence and characterization of cytoplasmic nuclear protein import factor p97. 1995, Pubmed
Corbett, Rna1p, a Ran/TC4 GTPase activating protein, is required for nuclear import. 1995, Pubmed
Coutavas, Characterization of proteins that interact with the cell-cycle regulatory protein Ran/TC4. 1993, Pubmed , Xenbase
Davis, Identification and characterization of a nuclear pore complex protein. 1986, Pubmed
Davis, The nuclear pore complex. 1995, Pubmed , Xenbase
Dingwall, A polypeptide domain that specifies migration of nucleoplasmin into the nucleus. 1982, Pubmed , Xenbase
Drivas, Characterization of four novel ras-like genes expressed in a human teratocarcinoma cell line. 1990, Pubmed
Fabre, Nuclear transport. 1994, Pubmed
Feldherr, Movement of a karyophilic protein through the nuclear pores of oocytes. 1984, Pubmed , Xenbase
Floer, The nuclear transport factor karyopherin beta binds stoichiometrically to Ran-GTP and inhibits the Ran GTPase activating protein. 1996, Pubmed
Forrester, Defects in mRNA 3'-end formation, transcription initiation, and mRNA transport associated with the yeast mutation prp20: possible coupling of mRNA processing and chromatin structure. 1992, Pubmed
Görlich, Isolation of a protein that is essential for the first step of nuclear protein import. 1994, Pubmed , Xenbase
Görlich, Two different subunits of importin cooperate to recognize nuclear localization signals and bind them to the nuclear envelope. 1995, Pubmed , Xenbase
Görlich, Distinct functions for the two importin subunits in nuclear protein import. 1995, Pubmed , Xenbase
Görlich, Nucleocytoplasmic transport. 1996, Pubmed
Görlich, A 41 amino acid motif in importin-alpha confers binding to importin-beta and hence transit into the nucleus. 1996, Pubmed , Xenbase
Hopper, The yeast RNA1 gene product necessary for RNA processing is located in the cytosol and apparently excluded from the nucleus. 1990, Pubmed
Imamoto, The nuclear pore-targeting complex binds to nuclear pores after association with a karyophile. 1995, Pubmed
Imamoto, In vivo evidence for involvement of a 58 kDa component of nuclear pore-targeting complex in nuclear protein import. 1995, Pubmed , Xenbase
Jarnik, Toward a more complete 3-D structure of the nuclear pore complex. 1991, Pubmed , Xenbase
Kadowaki, A conditional yeast mutant deficient in mRNA transport from nucleus to cytoplasm. 1992, Pubmed
Kalderon, Sequence requirements for nuclear location of simian virus 40 large-T antigen. , Pubmed
Klebe, Functional expression in Escherichia coli of the mitotic regulator proteins p24ran and p45rcc1 and fluorescence measurements of their interaction. 1993, Pubmed
Klebe, Interaction of the nuclear GTP-binding protein Ran with its regulatory proteins RCC1 and RanGAP1. 1995, Pubmed , Xenbase
Koepp, Dynamic localization of the nuclear import receptor and its interactions with transport factors. 1996, Pubmed
Kraemer, The essential yeast nucleoporin NUP159 is located on the cytoplasmic side of the nuclear pore complex and serves in karyopherin-mediated binding of transport substrate. 1995, Pubmed
Lounsbury, A family of proteins that stabilize the Ran/TC4 GTPase in its GTP-bound conformation. 1994, Pubmed
Lounsbury, Ran binding domains promote the interaction of Ran with p97/beta-karyopherin, linking the docking and translocation steps of nuclear import. 1996, Pubmed
Melchior, A functional homologue of the RNA1 gene product in Schizosaccharomyces pombe: purification, biochemical characterization, and identification of a leucine-rich repeat motif. 1993, Pubmed
Melchior, Inhibition of nuclear protein import by nonhydrolyzable analogues of GTP and identification of the small GTPase Ran/TC4 as an essential transport factor. 1993, Pubmed
Melchior, GTP hydrolysis by Ran occurs at the nuclear pore complex in an early step of protein import. 1995, Pubmed
Moore, The two steps of nuclear import, targeting to the nuclear envelope and translocation through the nuclear pore, require different cytosolic factors. 1992, Pubmed , Xenbase
Moore, The GTP-binding protein Ran/TC4 is required for protein import into the nucleus. 1993, Pubmed , Xenbase
Moore, Purification of a Ran-interacting protein that is required for protein import into the nucleus. 1994, Pubmed , Xenbase
Moroianu, Mammalian karyopherin alpha 1 beta and alpha 2 beta heterodimers: alpha 1 or alpha 2 subunit binds nuclear localization signal and beta subunit interacts with peptide repeat-containing nucleoporins. 1995, Pubmed
Nehrbass, Role of the nuclear transport factor p10 in nuclear import. 1996, Pubmed
Newmeyer, Assembly in vitro of nuclei active in nuclear protein transport: ATP is required for nucleoplasmin accumulation. 1986, Pubmed , Xenbase
Newmeyer, Nuclear import can be separated into distinct steps in vitro: nuclear pore binding and translocation. 1988, Pubmed , Xenbase
Ohtsubo, The RCC1 protein, a regulator for the onset of chromosome condensation locates in the nucleus and binds to DNA. 1989, Pubmed
Palacios, RAN/TC4 mutants identify a common requirement for snRNP and protein import into the nucleus. 1996, Pubmed , Xenbase
Panté, Toward the molecular dissection of protein import into nuclei. 1996, Pubmed
Panté, Interactions and three-dimensional localization of a group of nuclear pore complex proteins. 1994, Pubmed , Xenbase
Paschal, Identification of NTF2, a cytosolic factor for nuclear import that interacts with nuclear pore complex protein p62. 1995, Pubmed
Radu, Identification of a protein complex that is required for nuclear protein import and mediates docking of import substrate to distinct nucleoporins. 1995, Pubmed , Xenbase
Radu, The peptide repeat domain of nucleoporin Nup98 functions as a docking site in transport across the nuclear pore complex. 1995, Pubmed
Rexach, Protein import into nuclei: association and dissociation reactions involving transport substrate, transport factors, and nucleoporins. 1995, Pubmed
Richards, The C terminus of the nuclear RAN/TC4 GTPase stabilizes the GDP-bound state and mediates interactions with RCC1, RAN-GAP, and HTF9A/RANBP1. 1995, Pubmed
Richardson, Nuclear protein migration involves two steps: rapid binding at the nuclear envelope followed by slower translocation through nuclear pores. 1988, Pubmed , Xenbase
Schlenstedt, The GTP-bound form of the yeast Ran/TC4 homologue blocks nuclear protein import and appearance of poly(A)+ RNA in the cytoplasm. 1995, Pubmed
Schlenstedt, Mutants in a yeast Ran binding protein are defective in nuclear transport. 1995, Pubmed
Slot, A new method of preparing gold probes for multiple-labeling cytochemistry. 1985, Pubmed
Sweet, A GTPase distinct from Ran is involved in nuclear protein import. 1996, Pubmed
Tachibana, Loss of RCC1 leads to suppression of nuclear protein import in living cells. 1994, Pubmed
Weis, Identification of hSRP1 alpha as a functional receptor for nuclear localization sequences. 1995, Pubmed
Weis, The conserved amino-terminal domain of hSRP1 alpha is essential for nuclear protein import. 1996, Pubmed
Wu, Nup358, a cytoplasmically exposed nucleoporin with peptide repeats, Ran-GTP binding sites, zinc fingers, a cyclophilin A homologous domain, and a leucine-rich region. 1995, Pubmed
Yokoyama, A giant nucleopore protein that binds Ran/TC4. 1995, Pubmed