Note to users. If you're seeing this message, it means that your browser cannot find this page's style/presentation instructions -- or possibly that you are using a browser that does not support current Web standards. Find out more about why this message is appearing, and what you can do to make your experience of our site the best it can be.

Science 20 June 1997:
Vol. 276. no. 5320, pp. 1842 - 1844
DOI: 10.1126/science.276.5320.1842
Prev | Table of Contents | Next

Reports

Requirement of Guanosine Triphosphate-Bound Ran for Signal-Mediated Nuclear Protein Export

Stephanie A. Richards, * Kimberly L. Carey, Ian G. Macara dagger

A leucine-rich nuclear export signal (NES) allows rapid export of proteins from cell nuclei. Microinjection studies revealed a role for the guanosine triphosphatase (GTPase) Ran in NES-mediated export. Nuclear injection of a Ran mutant (Thr24 rightarrow  Asn) blocked protein export but not import, whereas depletion of the Ran nucleotide exchange factor RCC1 blocked protein import but not export. However, injection of Ran GTPase-activating protein (RanGAP) into RCC1-depleted cell nuclei inhibited export. Coinjection with Ran mutants insensitive to RanGAP prevented this inhibition. Therefore, NES-mediated protein export appears to require a Ran-GTP complex but does not require Ran-dependent GTP hydrolysis.

S. A. Richards and K. L. Carey, Department of Pathology, University of Vermont, Burlington, VT 05405, USA.
I. G. Macara, Center for Cell Signaling, Box 577, Health Sciences Center, University of Virginia, Charlottesville, VA 22908, USA.
*   Present address: Department of Cell Biology, Harvard Medical School, Boston, MA 02115, USA.

dagger    To whom correspondence should be addressed. E-mail: igm9c{at}virginia.edu

Read the Full Text


THIS ARTICLE HAS BEEN CITED BY OTHER ARTICLES:
Influence of heart failure on nucleocytoplasmic transport in human cardiomyocytes.
R. Cortes, E. Rosello-Lleti, M. Rivera, L. Martinez-Dolz, A. Salvador, I. Azorin, and M. Portoles (2009)
Cardiovasc Res
   Abstract »    Full Text »    PDF »
eEF1A Is a Novel Component of the Mammalian Nuclear Protein Export Machinery.
M. Khacho, K. Mekhail, K. Pilon-Larose, A. Pause, J. Cote, and S. Lee (2008)
Mol. Biol. Cell 19, 5296-5308
   Abstract »    Full Text »    PDF »
The Nup358-RanGAP Complex Is Required for Efficient Importin {alpha}/{beta}-dependent Nuclear Import.
S. Hutten, A. Flotho, F. Melchior, and R. H. Kehlenbach (2008)
Mol. Biol. Cell 19, 2300-2310
   Abstract »    Full Text »    PDF »
Activation of the DNA-dependent Protein Kinase Stimulates Nuclear Export of the Androgen Receptor in Vitro.
L. C. Shank, J. B. Kelley, D. Gioeli, C.-S. Yang, A. Spencer, L. A. Allison, and B. M. Paschal (2008)
J. Biol. Chem. 283, 10568-10580
   Abstract »    Full Text »    PDF »
Calcium-Dependent Regulation of NEMO Nuclear Export in Response to Genotoxic Stimuli.
C. M. Berchtold, Z.-H. Wu, T. T. Huang, and S. Miyamoto (2007)
Mol. Cell. Biol. 27, 497-509
   Abstract »    Full Text »    PDF »
Regulated nucleo-cytoplasmic shuttling of human aci-reductone dioxygenase (hADI1) and its potential role in mRNA processing..
I. Gotoh, T. Uekita, and M. Seiki (2007)
Genes Cells 12, 105-117
   Abstract »    Full Text »    PDF »
Systematic Deletion and Mitotic Localization of the Nuclear Pore Complex Proteins of Aspergillus nidulans.
A. H. Osmani, J. Davies, H.-L. Liu, A. Nile, and S. A. Osmani (2006)
Mol. Biol. Cell 17, 4946-4961
   Abstract »    Full Text »    PDF »
Nup214-Nup88 Nucleoporin Subcomplex Is Required for CRM1-mediated 60 S Preribosomal Nuclear Export.
R. Bernad, D. Engelsma, H. Sanderson, H. Pickersgill, and M. Fornerod (2006)
J. Biol. Chem. 281, 19378-19386
   Abstract »    Full Text »    PDF »
In vitro nuclear import.
M. Leslie (2006)
J. Cell Biol. 172, 789
   Full Text »    PDF »
Nuclear Export of the Nonenveloped Parvovirus Virion Is Directed by an Unordered Protein Signal Exposed on the Capsid Surface.
B. Maroto, N. Valle, R. Saffrich, and J. M. Almendral (2004)
J. Virol. 78, 10685-10694
   Abstract »    Full Text »    PDF »
Impairment of IL-12-Dependent STAT4 Nuclear Translocation in a Patient with Recurrent Mycobacterium avium Infection.
H. Toyoda, M. Ido, T. Hayashi, E. C. Gabazza, K. Suzuki, J. Bu, S. Tanaka, T. Nakano, H. Kamiya, J. Chipeta, et al. (2004)
J. Immunol. 172, 3905-3912
   Abstract »    Full Text »    PDF »
Cellular Requirements for CRM1 Import and Export.
X. Zhang, M. Yamada, N. Mabuchi, and H. Shida (2003)
J. Biochem. 134, 759-764
   Abstract »    Full Text »    PDF »
A Nuclear Kinesin-Like Protein Interacts with and Stimulates the Activity of the Leucine-Rich Nuclear Export Signal of the Human Immunodeficiency Virus Type 1 Rev Protein.
L. K. Venkatesh, T. Gettemeier, and G. Chinnadurai (2003)
J. Virol. 77, 7236-7243
   Abstract »    Full Text »    PDF »
Ran GTPase Cycle and Importins alpha and beta Are Essential for Spindle Formation and Nuclear Envelope Assembly in Living Caenorhabditis elegans Embryos.
P. Askjaer, V. Galy, E. Hannak, and I. W. Mattaj (2002)
Mol. Biol. Cell 13, 4355-4370
   Abstract »    Full Text »    PDF »
Reconstitution of nuclear protein export in isolated nuclear envelopes.
J. P. Siebrasse, E. Coutavas, and R. Peters (2002)
J. Cell Biol. 158, 849-854
   Abstract »    Full Text »    PDF »
Stat5B Shuttles Between Cytoplasm and Nucleus in a Cytokine-Dependent and -Independent Manner.
R. Zeng, Y. Aoki, M. Yoshida, K.-i. Arai, and S. Watanabe (2002)
J. Immunol. 168, 4567-4575
   Abstract »    Full Text »    PDF »
Systems Analysis of Ran Transport.
A. E. Smith, B. M. Slepchenko, J. C. Schaff, L. M. Loew, and I. G. Macara (2002)
Science 295, 488-491
   Abstract »    Full Text »    PDF »
Transport into and out of the Nucleus.
I. G. Macara (2001)
Microbiol. Mol. Biol. Rev. 65, 570-594
   Abstract »    Full Text »    PDF »
The Docking of Kinesins, KIF5B and KIF5C, to Ran-binding Protein 2 (RanBP2) Is Mediated via a Novel RanBP2 Domain.
Y. Cai, B. B. Singh, A. Aslanukov, H. Zhao, and P. A. Ferreira (2001)
J. Biol. Chem. 276, 41594-41602
   Abstract »    Full Text »    PDF »
In Vitro Analysis of Nuclear Transport Mediated by the C-terminal Shuttle Domain of Tap.
I. Schmitt and L. Gerace (2001)
J. Biol. Chem. 276, 42355-42363
   Abstract »    Full Text »    PDF »
Ran-Binding Protein 3 Is a Cofactor for Crm1-Mediated Nuclear Protein Export.
M. E. Lindsay, J. M. Holaska, K. Welch, B. M. Paschal, and I. G. Macara (2001)
J. Cell Biol. 153, 1391-1402
   Abstract »    Full Text »    PDF »
Inhibition of Nuclear Import by Protein Kinase B (Akt) Regulates the Subcellular Distribution and Activity of the Forkhead Transcription Factor AFX.
A. M. Brownawell, G. J. P. L. Kops, I. G. Macara, and B. M. T. Burgering (2001)
Mol. Cell. Biol. 21, 3534-3546
   Abstract »    Full Text »
A Single Amino Acid Substitution in the ICP27 Protein of Herpes Simplex Virus Type 1 Is Responsible for Its Resistance to Leptomycin B.
T. Murata, F. Goshima, T. Koshizuka, H. Takakuwa, and Y. Nishiyama (2001)
J. Virol. 75, 1039-1043
   Abstract »    Full Text »
Calreticulin Is a Receptor for Nuclear Export.
J. M. Holaska, B. E. Black, D. C. Love, J. A. Hanover, J. Leszyk, and B. M. Paschal (2001)
J. Cell Biol. 152, 127-140
   Abstract »    Full Text »    PDF »
RanGTP-Binding Protein NXT1 Facilitates Nuclear Export of Different Classes of RNA In Vitro.
B. Ossareh-Nazari, C. Maison, B. E. Black, L. Lévesque, B. M. Paschal, and C. Dargemont (2000)
Mol. Cell. Biol. 20, 4562-4571
   Abstract »    Full Text »
Yeast Ran-Binding Protein 1 (Yrb1) Shuttles between the Nucleus and Cytoplasm and Is Exported from the Nucleus via a CRM1 (XPO1)-Dependent Pathway.
M. Künzler, T. Gerstberger, F. Stutz, F. R. Bischoff, and E. Hurt (2000)
Mol. Cell. Biol. 20, 4295-4308
   Abstract »    Full Text »
Facilitated Nucleocytoplasmic Shuttling of the Ran Binding Protein RanBP1.
K. Plafker and I. G. Macara (2000)
Mol. Cell. Biol. 20, 3510-3521
   Abstract »    Full Text »
Nuclear Import of the Ran Exchange Factor, Rcc1, Is Mediated by at Least Two Distinct Mechanisms.
M. E. Nemergut and I. G. Macara (2000)
J. Cell Biol. 149, 835-850
   Abstract »    Full Text »    PDF »
Nuclear Import of Ikappa Balpha Is Accomplished by a Ran-Independent Transport Pathway.
S. Sachdev, S. Bagchi, D. D. Zhang, A. C. Mings, and M. Hannink (2000)
Mol. Cell. Biol. 20, 1571-1582
   Abstract »    Full Text »
Random Mutagenesis and Functional Analysis of the Ran-binding Protein, RanBP1.
C. Petersen, N. Orem, J. Trueheart, J. W. Thorner, and I. G. Macara (2000)
J. Biol. Chem. 275, 4081-4091
   Abstract »    Full Text »    PDF »
Nucleocytoplasmic shuttling: a novel in vivo property of antisense phosphorothioate oligodeoxynucleotides.
P. Lorenz, T. Misteli, B. F. Baker, C. F. Bennett, and D. L. Spector (2000)
Nucleic Acids Res. 28, 582-592
   Abstract »    Full Text »    PDF »
The ran decathlon: multiple roles of Ran.
S Sazer and M Dasso (2000)
J. Cell Sci. 113, 1111-1118
   Abstract »    PDF »
The Zinc Finger Cluster Domain of RanBP2 Is a Specific Docking Site for the Nuclear Export Factor, Exportin-1.
B. B. Singh, H. H. Patel, R. Roepman, D. Schick, and P. A. Ferreira (1999)
J. Biol. Chem. 274, 37370-37378
   Abstract »    Full Text »    PDF »
RanBP3 Contains an Unusual Nuclear Localization Signal That Is Imported Preferentially by Importin-alpha 3.
K. Welch, J. Franke, M. Kohler, and I. G. Macara (1999)
Mol. Cell. Biol. 19, 8400-8411
   Abstract »    Full Text »    PDF »
Identification of an NTF2-Related Factor That Binds Ran-GTP and Regulates Nuclear Protein Export.
B. E. Black, L. Levesque, J. M. Holaska, T. C. Wood, and B. M. Paschal (1999)
Mol. Cell. Biol. 19, 8616-8624
   Abstract »    Full Text »    PDF »
Kap104p-mediated Nuclear Import. NUCLEAR LOCALIZATION SIGNALS IN mRNA-BINDING PROTEINS AND THE ROLE OF RAN AND RNA.
D. C. Y. Lee and J. D. Aitchison (1999)
J. Biol. Chem. 274, 29031-29037
   Abstract »    Full Text »    PDF »
Beyond Nuclear Transport: Ran-Gtp as a Determinant of Spindle Assembly.
J. A. Kahana and D. W. Cleveland (1999)
J. Cell Biol. 146, 1205-1210
   Full Text »    PDF »
The direction of transport through the nuclear pore can be inverted.
M. V. Nachury and K. Weis (1999)
PNAS 96, 9622-9627
   Abstract »    Full Text »    PDF »
Association with the Cellular Export Receptor CRM 1 Mediates Function and Intracellular Localization of Epstein-Barr Virus SM Protein, a Regulator of Gene Expression.
S. M. Boyle, V. Ruvolo, A. K. Gupta, and S. Swaminathan (1999)
J. Virol. 73, 6872-6881
   Abstract »    Full Text »
Putative Reaction Intermediates in Crm1-mediated Nuclear Protein Export.
M. Floer and G. Blobel (1999)
J. Biol. Chem. 274, 16279-16286
   Abstract »    Full Text »    PDF »
A Role for RanBP1 in the Release of CRM1 from the Nuclear Pore Complex in a Terminal Step of Nuclear Export.
R. H. Kehlenbach, A. Dickmanns, A. Kehlenbach, T. Guan, and L. Gerace (1999)
J. Cell Biol. 145, 645-657
   Abstract »    Full Text »    PDF »
RAE1 Is a Shuttling mRNA Export Factor That Binds to a GLEBS-like NUP98 Motif at the Nuclear Pore Complex through Multiple Domains.
C. E.J. Pritchard, M. Fornerod, L. H. Kasper, and J. M.A. van Deursen (1999)
J. Cell Biol. 145, 237-254
   Abstract »    Full Text »    PDF »
Characterization of Ikappa Balpha Nuclear Import Pathway.
P. Turpin, R. T. Hay, and C. Dargemont (1999)
J. Biol. Chem. 274, 6804-6812
   Abstract »    Full Text »    PDF »
A Monoclonal Antibody to the COOH-terminal Acidic Portion of Ran Inhibits Both the Recycling of Ran and Nuclear Protein Import in Living Cells.
M. Hieda, T. Tachibana, F. Yokoya, S. Kose, N. Imamoto, and Y. Yoneda (1999)
J. Cell Biol. 144, 645-655
   Abstract »    Full Text »    PDF »
beta -Subunit of Nuclear Pore-targeting Complex (Importin-beta ) Can Be Exported from the Nucleus in a Ran-independent Manner.
S. Kose, N. Imamoto, T. Tachibana, M. Yoshida, and Y. Yoneda (1999)
J. Biol. Chem. 274, 3946-3952
   Abstract »    Full Text »    PDF »
Importin beta  Can Mediate the Nuclear Import of an Arginine-Rich Nuclear Localization Signal in the Absence of Importin alpha.
D. Palmeri and M. H. Malim (1999)
Mol. Cell. Biol. 19, 1218-1225
   Abstract »    Full Text »    PDF »
Antagonistic effects of NES and NLS motifs determine S. cerevisiae Rna1p subcellular distribution.
W Feng, A. Benko, J. Lee, D. Stanford, and A. Hopper (1999)
J. Cell Sci. 112, 339-347
   Abstract »    PDF »
The Specificity of the CRM1-Rev Nuclear Export Signal Interaction Is Mediated by RanGTP.
P. Askjaer, T. H. Jensen, J. Nilsson, L. Englmeier, and J. Kjems (1998)
J. Biol. Chem. 273, 33414-33422
   Abstract »    Full Text »    PDF »
Proofreading and Aminoacylation of tRNAs Before Export from the Nucleus.
E. Lund and J. E. Dahlberg (1998)
Science 282, 2082-2085
   Abstract »    Full Text »
A cytosolic activity distinct from Crm1 mediates nuclear export of protein kinase inhibitor in permeabilized cells.
J. M. Holaska and B. M. Paschal (1998)
PNAS 95, 14739-14744
   Abstract »    Full Text »    PDF »
Yeast Los1p Has Properties of an Exportin-Like Nucleocytoplasmic Transport Factor for tRNA.
K. Hellmuth, D. M. Lau, F. R. Bischoff, M. Künzler, E. Hurt, and G. Simos (1998)
Mol. Cell. Biol. 18, 6374-6386
   Abstract »    Full Text »
Nuclear mRNA Export Requires Complex Formation between Mex67p and Mtr2p at the Nuclear Pores.
H. Santos-Rosa, H. Moreno, G. Simos, A. Segref, B. Fahrenkrog, N. Panté, and E. Hurt (1998)
Mol. Cell. Biol. 18, 6826-6838
   Abstract »    Full Text »
Mechanisms of Progesterone Receptor Export from Nuclei: Role of Nuclear Localization Signal, Nuclear Export Signal, and Ran Guanosine Triphosphate.
R. K. Tyagi, L. Amazit, P. Lescop, E. Milgrom, and A. Guiochon-Mantel (1998)
Mol. Endocrinol. 12, 1684-1695
   Abstract »    Full Text »
Nuclear RNA export.
F. Stutz and M. Rosbash (1998)
Genes & Dev. 12, 3303-3319
   Full Text »
Ran and Nuclear Transport.
M. S. Moore (1998)
J. Biol. Chem. 273, 22857-22860
   Full Text »    PDF »
Reconstitution of HIV-1 Rev nuclear export: Independent requirements for nuclear import and export.
D. C. Love, T. D. Sweitzer, and J. A. Hanover (1998)
PNAS 95, 10608-10613
   Abstract »    Full Text »    PDF »
Control of Cyclin B1 localization through regulated binding of the nuclear export factor CRM1.
J. Yang, E. S.G. Bardes, J. D. Moore, J. Brennan, M. A. Powers, and S. Kornbluth (1998)
Genes & Dev. 12, 2131-2143
   Abstract »    Full Text »
A member of the Ran-binding protein family, Yrb2p, is involved in nuclear protein export.
T. Taura, H. Krebber, and P. A. Silver (1998)
PNAS 95, 7427-7432
   Abstract »    Full Text »    PDF »
Nucleocytoplasmic Shuttling Factors Including Ran and CRM1 Mediate Nuclear Export of NFAT In Vitro.
R. H. Kehlenbach, A. Dickmanns, and L. Gerace (1998)
J. Cell Biol. 141, 863-874
   Abstract »    Full Text »    PDF »
The role of the ran GTPase in nuclear assembly and DNA replication: characterisation of the effects of Ran mutants.
M Hughes, C Zhang, J. Avis, C. Hutchison, and P. Clarke (1998)
J. Cell Sci. 111, 3017-3026
   Abstract »    PDF »
Nuclear import of protein kinase C occurs by a mechanism distinct from the mechanism used by proteins with a classical nuclear localization signal.
D Schmalz, F Hucho, and K Buchner (1998)
J. Cell Sci. 111, 1823-1830
   Abstract »    PDF »
Nucleocytoplasmic Recycling of the Nuclear Localization Signal Receptor {alpha} Subunit In Vivo Is Dependent on a Nuclear Export Signal, Energy, and RCC1.
I. Boche and E. Fanning (1997)
J. Cell Biol. 139, 313-325
   Abstract »    Full Text »    PDF »
Evidence for a Role of CRM1 in Signal-Mediated Nuclear Protein Export.
B. Ossareh-Nazari, F. Bachelerie, and C. Dargemont (1997)
Science 278, 141-144
   Abstract »    Full Text »
Selection and nuclear immobilization of exportable RNAs.
C. Grimm, E. Lund, and J. E. Dahlberg (1997)
PNAS 94, 10122-10127
   Abstract »    Full Text »    PDF »
Xenopus Ran-binding protein 1: molecular interactions and effects on nuclear assembly in Xenopus egg extracts.
F. Nicolas, C Zhang, M Hughes, M Goldberg, S Watton, and P Clarke (1997)
J. Cell Sci. 110, 3019-3030
   Abstract »    PDF »
Cytoplasmic, Nuclear, and Golgi Localization of RGS Proteins. EVIDENCE FOR N-TERMINAL AND RGS DOMAIN SEQUENCES AS INTRACELLULAR TARGETING MOTIFS.
T. K. Chatterjee and R. A. Fisher (2000)
J. Biol. Chem. 275, 24013-24021
   Abstract »    Full Text »    PDF »
Characterization of the Nuclear Import Pathway for HIV-1 Integrase.
C. Depienne, A. Mousnier, H. Leh, E. Le Rouzic, D. Dormont, S. Benichou, and C. Dargemont (2001)
J. Biol. Chem. 276, 18102-18107
   Abstract »    Full Text »    PDF »
Exportin-5, a novel karyopherin, mediates nuclear export of double-stranded RNA binding proteins.
A. M. Brownawell and I. G. Macara (2002)
J. Cell Biol. 156, 53-64
   Abstract »    Full Text »    PDF »


To Advertise     Find Products

Science. ISSN 0036-8075 (print), 1095-9203 (online)