PMCCPMCCPMCC

Search tips
Search criteria 

Advanced

 
Logo of jcellbiolHomeThe Rockefeller University PressEditorsContactInstructions for AuthorsThis issue
 
J Cell Biol. 1996 September 1; 134(5): 1157–1168.
PMCID: PMC2120988

A nuclear export signal is essential for the cytosolic localization of the Ran binding protein, RanBP1

Abstract

RanBP1 is a Ran/TC4 binding protein that can promote the interaction between Ran and beta-importin /beta-karyopherin, a component of the docking complex for nuclear protein cargo. This interaction occurs through a Ran binding domain (RBD). Here we show that RanBP1 is primarily cytoplasmic, but the isolated RBD accumulates in the nucleus. A region COOH-terminal to the RBD is responsible for this cytoplasmic localization. This domain acts heterologously, localizing a nuclear cyclin B1 mutant to the cytoplasm. The domain contains a nuclear export signal that is necessary but not sufficient for the nuclear export of a functional RBD In transiently transfected cells, epitope-tagged RanBP1 promotes dexamethasone-dependent nuclear accumulation of a glucocorticoid receptor-green fluorescent protein fusion, but the isolated RBD potently inhibits this accumulation. The cytosolic location of RanBP1 may therefore be important for nuclear protein import. RanBP1 may provide a key link between the nuclear import and export pathways.

Full Text

The Full Text of this article is available as a PDF (2.7M).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Aebi M, Clark MW, Vijayraghavan U, Abelson J. A yeast mutant, PRP20, altered in mRNA metabolism and maintenance of the nuclear structure, is defective in a gene homologous to the human gene RCC1 which is involved in the control of chromosome condensation. Mol Gen Genet. 1990 Oct;224(1):72–80. [PubMed]
  • Beddow AL, Richards SA, Orem NR, Macara IG. The Ran/TC4 GTPase-binding domain: identification by expression cloning and characterization of a conserved sequence motif. Proc Natl Acad Sci U S A. 1995 Apr 11;92(8):3328–3332. [PubMed]
  • Belhumeur P, Lee A, Tam R, DiPaolo T, Fortin N, Clark MW. GSP1 and GSP2, genetic suppressors of the prp20-1 mutant in Saccharomyces cerevisiae: GTP-binding proteins involved in the maintenance of nuclear organization. Mol Cell Biol. 1993 Apr;13(4):2152–2161. [PMC free article] [PubMed]
  • Bischoff FR, Ponstingl H. Mitotic regulator protein RCC1 is complexed with a nuclear ras-related polypeptide. Proc Natl Acad Sci U S A. 1991 Dec 1;88(23):10830–10834. [PubMed]
  • Bischoff FR, Ponstingl H. Catalysis of guanine nucleotide exchange on Ran by the mitotic regulator RCC1. Nature. 1991 Nov 7;354(6348):80–82. [PubMed]
  • Bischoff FR, Krebber H, Smirnova E, Dong W, Ponstingl H. Co-activation of RanGTPase and inhibition of GTP dissociation by Ran-GTP binding protein RanBP1. EMBO J. 1995 Feb 15;14(4):705–715. [PubMed]
  • Bogerd HP, Fridell RA, Madore S, Cullen BR. Identification of a novel cellular cofactor for the Rev/Rex class of retroviral regulatory proteins. Cell. 1995 Aug 11;82(3):485–494. [PubMed]
  • Bressan A, Somma MP, Lewis J, Santolamazza C, Copeland NG, Gilbert DJ, Jenkins NA, Lavia P. Characterization of the opposite-strand genes from the mouse bidirectionally transcribed HTF9 locus. Gene. 1991 Jul 22;103(2):201–209. [PubMed]
  • Clark KL, Sprague GF., Jr Yeast pheromone response pathway: characterization of a suppressor that restores mating to receptorless mutants. Mol Cell Biol. 1989 Jun;9(6):2682–2694. [PMC free article] [PubMed]
  • Clarke PR, Klebe C, Wittinghofer A, Karsenti E. Regulation of Cdc2/cyclin B activation by Ran, a Ras-related GTPase. J Cell Sci. 1995 Mar;108(Pt 3):1217–1225. [PubMed]
  • Coutavas E, Ren M, Oppenheim JD, D'Eustachio P, Rush MG. Characterization of proteins that interact with the cell-cycle regulatory protein Ran/TC4. Nature. 1993 Dec 9;366(6455):585–587. [PubMed]
  • Dasso M, Nishitani H, Kornbluth S, Nishimoto T, Newport JW. RCC1, a regulator of mitosis, is essential for DNA replication. Mol Cell Biol. 1992 Aug;12(8):3337–3345. [PMC free article] [PubMed]
  • Davis LI. The nuclear pore complex. Annu Rev Biochem. 1995;64:865–896. [PubMed]
  • Eckert RL, Katzenellenbogen BS. Physical properties of estrogen receptor complexes in MCF-7 human breast cancer cells. Differences with anti-estrogen and estrogen. J Biol Chem. 1982 Aug 10;257(15):8840–8846. [PubMed]
  • Fantozzi DA, Harootunian AT, Wen W, Taylor SS, Feramisco JR, Tsien RY, Meinkoth JL. Thermostable inhibitor of cAMP-dependent protein kinase enhances the rate of export of the kinase catalytic subunit from the nucleus. J Biol Chem. 1994 Jan 28;269(4):2676–2686. [PubMed]
  • Fischer U, Huber J, Boelens WC, Mattaj IW, Lührmann R. The HIV-1 Rev activation domain is a nuclear export signal that accesses an export pathway used by specific cellular RNAs. Cell. 1995 Aug 11;82(3):475–483. [PubMed]
  • Forrester W, Stutz F, Rosbash M, Wickens M. 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. Genes Dev. 1992 Oct;6(10):1914–1926. [PubMed]
  • Gerace L. Nuclear export signals and the fast track to the cytoplasm. Cell. 1995 Aug 11;82(3):341–344. [PubMed]
  • Guddat U, Bakken AH, Pieler T. Protein-mediated nuclear export of RNA: 5S rRNA containing small RNPs in xenopus oocytes. Cell. 1990 Feb 23;60(4):619–628. [PubMed]
  • Hayashi N, Yokoyama N, Seki T, Azuma Y, Ohba T, Nishimoto T. RanBP1, a Ras-like nuclear G protein binding to Ran/TC4, inhibits RCC1 via Ran/TC4. Mol Gen Genet. 1995 Jun 25;247(6):661–669. [PubMed]
  • Heim R, Cubitt AB, Tsien RY. Improved green fluorescence. Nature. 1995 Feb 23;373(6516):663–664. [PubMed]
  • Izaurralde E, Mattaj IW. RNA export. Cell. 1995 Apr 21;81(2):153–159. [PubMed]
  • Kadowaki T, Goldfarb D, Spitz LM, Tartakoff AM, Ohno M. Regulation of RNA processing and transport by a nuclear guanine nucleotide release protein and members of the Ras superfamily. EMBO J. 1993 Jul;12(7):2929–2937. [PubMed]
  • Kornbluth S, Dasso M, Newport J. Evidence for a dual role for TC4 protein in regulating nuclear structure and cell cycle progression. J Cell Biol. 1994 May;125(4):705–719. [PMC free article] [PubMed]
  • Lounsbury KM, Beddow AL, Macara IG. A family of proteins that stabilize the Ran/TC4 GTPase in its GTP-bound conformation. J Biol Chem. 1994 Apr 15;269(15):11285–11290. [PubMed]
  • Lounsbury KM, Richards SA, Perlungher RR, Macara IG. Ran binding domains promote the interaction of Ran with p97/beta-karyopherin, linking the docking and translocation steps of nuclear import. J Biol Chem. 1996 Feb 2;271(5):2357–2360. [PubMed]
  • Matsumoto T, Beach D. Premature initiation of mitosis in yeast lacking RCC1 or an interacting GTPase. Cell. 1991 Jul 26;66(2):347–360. [PubMed]
  • Mattingly RR, Sorisky A, Brann MR, Macara IG. Muscarinic receptors transform NIH 3T3 cells through a Ras-dependent signalling pathway inhibited by the Ras-GTPase-activating protein SH3 domain. Mol Cell Biol. 1994 Dec;14(12):7943–7952. [PMC free article] [PubMed]
  • Melchior F, Gerace L. Mechanisms of nuclear protein import. Curr Opin Cell Biol. 1995 Jun;7(3):310–318. [PubMed]
  • Melchior F, Paschal B, Evans J, Gerace L. Inhibition of nuclear protein import by nonhydrolyzable analogues of GTP and identification of the small GTPase Ran/TC4 as an essential transport factor. J Cell Biol. 1993 Dec;123(6 Pt 2):1649–1659. [PMC free article] [PubMed]
  • Melchior F, Guan T, Yokoyama N, Nishimoto T, Gerace L. GTP hydrolysis by Ran occurs at the nuclear pore complex in an early step of protein import. J Cell Biol. 1995 Nov;131(3):571–581. [PMC free article] [PubMed]
  • Michael WM, Choi M, Dreyfuss G. A nuclear export signal in hnRNP A1: a signal-mediated, temperature-dependent nuclear protein export pathway. Cell. 1995 Nov 3;83(3):415–422. [PubMed]
  • Moore MS, Blobel G. The GTP-binding protein Ran/TC4 is required for protein import into the nucleus. Nature. 1993 Oct 14;365(6447):661–663. [PubMed]
  • Moroianu J, Blobel G. Protein export from the nucleus requires the GTPase Ran and GTP hydrolysis. Proc Natl Acad Sci U S A. 1995 May 9;92(10):4318–4322. [PubMed]
  • Ouspenski II, Mueller UW, Matynia A, Sazer S, Elledge SJ, Brinkley BR. Ran-binding protein-1 is an essential component of the Ran/RCC1 molecular switch system in budding yeast. J Biol Chem. 1995 Feb 3;270(5):1975–1978. [PubMed]
  • Pines J, Hunter T. The differential localization of human cyclins A and B is due to a cytoplasmic retention signal in cyclin B. EMBO J. 1994 Aug 15;13(16):3772–3781. [PubMed]
  • Piñol-Roma S, Dreyfuss G. Shuttling of pre-mRNA binding proteins between nucleus and cytoplasm. Nature. 1992 Feb 20;355(6362):730–732. [PubMed]
  • Ren M, Coutavas E, D'Eustachio P, Rush MG. Effects of mutant Ran/TC4 proteins on cell cycle progression. Mol Cell Biol. 1994 Jun;14(6):4216–4224. [PMC free article] [PubMed]
  • Richards SA, Lounsbury KM, Macara IG. The C terminus of the nuclear RAN/TC4 GTPase stabilizes the GDP-bound state and mediates interactions with RCC1, RAN-GAP, and HTF9A/RANBP1. J Biol Chem. 1995 Jun 16;270(24):14405–14411. [PubMed]
  • Rudt F, Pieler T. Cytoplasmic retention and nuclear import of 5S ribosomal RNA containing RNPs. EMBO J. 1996 Mar 15;15(6):1383–1391. [PubMed]
  • Schlenstedt G, Saavedra C, Loeb JD, Cole CN, Silver PA. The GTP-bound form of the yeast Ran/TC4 homologue blocks nuclear protein import and appearance of poly(A)+ RNA in the cytoplasm. Proc Natl Acad Sci U S A. 1995 Jan 3;92(1):225–229. [PubMed]
  • Schlenstedt G, Wong DH, Koepp DM, Silver PA. Mutants in a yeast Ran binding protein are defective in nuclear transport. EMBO J. 1995 Nov 1;14(21):5367–5378. [PubMed]
  • Stutz F, Neville M, Rosbash M. Identification of a novel nuclear pore-associated protein as a functional target of the HIV-1 Rev protein in yeast. Cell. 1995 Aug 11;82(3):495–506. [PubMed]
  • Wen W, Meinkoth JL, Tsien RY, Taylor SS. Identification of a signal for rapid export of proteins from the nucleus. Cell. 1995 Aug 11;82(3):463–473. [PubMed]
  • Wu J, Matunis MJ, Kraemer D, Blobel G, Coutavas E. Nup358, a cytoplasmically exposed nucleoporin with peptide repeats, Ran-GTP binding sites, zinc fingers, a cyclophilin A homologous domain, and a leucine-rich region. J Biol Chem. 1995 Jun 9;270(23):14209–14213. [PubMed]
  • Yokoyama N, Hayashi N, Seki T, Panté N, Ohba T, Nishii K, Kuma K, Hayashida T, Miyata T, Aebi U, et al. A giant nucleopore protein that binds Ran/TC4. Nature. 1995 Jul 13;376(6536):184–188. [PubMed]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press