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.
AAAS Promotion

Site Tools

  • AAAS
  • Subscribe
  • Feedback

Site Search

Search Advanced

Science 29 March 2002:
Vol. 295. no. 5564, pp. 2452 - 2456
DOI: 10.1126/science.1068798
Prev | Table of Contents | Next

Reports

Visualization of a Ran-GTP Gradient in Interphase and Mitotic Xenopus Egg Extracts

Petr Kalab, Karsten Weis,* Rebecca Heald*

The small guanosine triphosphatase Ran is loaded with guanosine triphosphate (GTP) by the chromatin-bound guanine nucleotide exchange factor RCC1 and releases import cargoes in the nucleus during interphase. In mitosis, Ran-GTP promotes spindle assembly around chromosomes by locally discharging cargoes that regulate microtubule dynamics and organization. We used fluorescence resonance energy transfer-based biosensors to visualize gradients of Ran-GTP and liberated cargoes around chromosomes in mitotic Xenopus egg extracts. Both gradients were required to assemble and maintain spindle structure. During interphase, Ran-GTP was highly enriched in the nucleoplasm, and a steep concentration difference between nuclear and cytoplasmic Ran-GTP was established, providing evidence for a Ran-GTP gradient surrounding chromosomes throughout the cell cycle.

Department of Molecular and Cell Biology, University of California, Berkeley, CA 94720-3200, USA.
*   To whom correspondence should be addressed. E-mail: heald{at}socrates.berkeley.edu (R.H.); kweis{at}uclink4.berkeley.edu (K.W.)

Read the Full Text


THIS ARTICLE HAS BEEN CITED BY OTHER ARTICLES:
Regulation of Microtubule Assembly and Organization in Mitosis by the AAA+ ATPase Pontin.
D. Ducat, S.-i. Kawaguchi, H. Liu, J. R. Yates III, and Y. Zheng (2008)
Mol. Biol. Cell 19, 3097-3110
   Abstract »    Full Text »    PDF »
Two Distinct Interacting Classes of Nuclear Envelope-Associated Coiled-Coil Proteins Are Required for the Tissue-Specific Nuclear Envelope Targeting of Arabidopsis RanGAP.
Q. Zhao, J. Brkljacic, and I. Meier (2008)
PLANT CELL 20, 1639-1651
   Abstract »    Full Text »    PDF »
RanBP10 Is a Cytoplasmic Guanine Nucleotide Exchange Factor That Modulates Noncentrosomal Microtubules.
H. Schulze, M. Dose, M. Korpal, I. Meyer, J. E. Italiano Jr., and R. A. Shivdasani (2008)
J. Biol. Chem. 283, 14109-14119
   Abstract »    Full Text »    PDF »
The RanGTP gradient - a GPS for the mitotic spindle.
P. Kalab and R. Heald (2008)
J. Cell Sci. 121, 1577-1586
   Abstract »    Full Text »    PDF »
Cdk11 is a RanGTP-dependent microtubule stabilization factor that regulates spindle assembly rate.
H. Yokoyama, O. J. Gruss, S. Rybina, M. Caudron, M. Schelder, M. Wilm, I. W. Mattaj, and E. Karsenti (2008)
J. Cell Biol. 180, 867-875
   Abstract »    Full Text »    PDF »
Importin-{beta} and the small guanosine triphosphatase Ran mediate chromosome loading of the human chromokinesin Kid.
K. Tahara, M. Takagi, M. Ohsugi, T. Sone, F. Nishiumi, K. Maeshima, Y. Horiuchi, N. Tokai-Nishizumi, F. Imamoto, T. Yamamoto, et al. (2008)
J. Cell Biol. 180, 493-506
   Abstract »    Full Text »    PDF »
Toward a High-Resolution View of Nuclear Dynamics.
L. Trinkle-Mulcahy and A. I. Lamond (2007)
Science 318, 1402-1407
   Abstract »    Full Text »    PDF »
Hyperosmotic Stress Signaling to the Nucleus Disrupts the Ran Gradient and the Production of RanGTP.
J. B. Kelley and B. M. Paschal (2007)
Mol. Biol. Cell 18, 4365-4376
   Abstract »    Full Text »    PDF »
RCC1 isoforms differ in their affinity for chromatin, molecular interactions and regulation by phosphorylation.
F. E. Hood and P. R. Clarke (2007)
J. Cell Sci. 120, 3436-3445
   Abstract »    Full Text »    PDF »
Cooperative mechanisms of mitotic spindle formation.
C. B. O'Connell and A. L. Khodjakov (2007)
J. Cell Sci. 120, 1717-1722
   Abstract »    Full Text »    PDF »
The Karyopherin Kap95 Regulates Nuclear Pore Complex Assembly into Intact Nuclear Envelopes In Vivo.
K. J. Ryan, Y. Zhou, and S. R. Wente (2007)
Mol. Biol. Cell 18, 886-898
   Abstract »    Full Text »    PDF »
Classical Nuclear Localization Signals: Definition, Function, and Interaction with Importin {alpha}.
A. Lange, R. E. Mills, C. J. Lange, M. Stewart, S. E. Devine, and A. H. Corbett (2007)
J. Biol. Chem. 282, 5101-5105
   Abstract »    Full Text »    PDF »
A centriole- and RanGTP-independent spindle assembly pathway in meiosis I of vertebrate oocytes.
J. Dumont, S. Petri, F. Pellegrin, M.-E. Terret, M. T. Bohnsack, P. Rassinier, V. Georget, P. Kalab, O. J. Gruss, and M.-H. Verlhac (2007)
J. Cell Biol. 176, 295-305
   Abstract »    Full Text »    PDF »
Phosphorylation of Maskin by Aurora-A Is Regulated by RanGTP and Importin beta.
A. J. Albee, W. Tao, and C. Wiese (2006)
J. Biol. Chem. 281, 38293-38301
   Abstract »    Full Text »    PDF »
The Nup107-160 Nucleoporin Complex Is Required for Correct Bipolar Spindle Assembly.
A. V. Orjalo, A. Arnaoutov, Z. Shen, Y. Boyarchuk, S. G. Zeitlin, B. Fontoura, S. Briggs, M. Dasso, and D. J. Forbes (2006)
Mol. Biol. Cell 17, 3806-3818
   Abstract »    Full Text »    PDF »
A picornavirus protein interacts with Ran-GTPase and disrupts nucleocytoplasmic transport.
F. W. Porter, Y. A. Bochkov, A. J. Albee, C. Wiese, and A. C. Palmenberg (2006)
PNAS 103, 12417-12422
   Abstract »    Full Text »    PDF »
Nuclear RanGAP Is Required for the Heterochromatin Assembly and Is Reciprocally Regulated by Histone H3 and Clr4 Histone Methyltransferase in Schizosaccharomyces pombe.
H. Nishijima, J.-i. Nakayama, T. Yoshioka, A. Kusano, H. Nishitani, K.-i. Shibahara, and T. Nishimoto (2006)
Mol. Biol. Cell 17, 2524-2536
   Abstract »    Full Text »    PDF »
Kap120 Functions as a Nuclear Import Receptor for Ribosome Assembly Factor Rpf1 in Yeast.
S. Caesar, M. Greiner, and G. Schlenstedt (2006)
Mol. Cell. Biol. 26, 3170-3180
   Abstract »    Full Text »    PDF »
Ran Is Required before Metaphase for Spindle Assembly and Chromosome Alignment and after Metaphase for Chromosome Segregation and Spindle Midbody Organization.
R. V. Silverman-Gavrila and A. Wilde (2006)
Mol. Biol. Cell 17, 2069-2080
   Abstract »    Full Text »    PDF »
In vitro nuclear import.
M. Leslie (2006)
J. Cell Biol. 172, 789
   Full Text »    PDF »
Loss of RanGEF/Pim1 activity abolishes the orchestration of Ran-mediated mitotic cellular events in S. pombe.
E. Hirose, M. Mukai, A. Shimada, H. Nishitani, Y. Shibata, and T. Nishimoto (2006)
Genes Cells 11, 29-46
   Abstract »    Full Text »    PDF »
Anaphase-Promoting Complex/Cyclosome Controls the Stability of TPX2 during Mitotic Exit.
S. Stewart and G. Fang (2005)
Mol. Cell. Biol. 25, 10516-10527
   Abstract »    Full Text »    PDF »
Cytoskeleton and cell cycle control during meiotic maturation of the mouse oocyte: integrating time and space.
S. Brunet and B. Maro (2005)
Reproduction 130, 801-811
   Abstract »    Full Text »    PDF »
The Fission Yeast Schizosaccharomyces pombe Has Two Importin-{alpha} Proteins, Imp1p and Cut15p, Which Have Common and Unique Functions in Nucleocytoplasmic Transport and Cell Cycle Progression.
M. Umeda, S. Izaddoost, I. Cushman, M. S. Moore, and S. Sazer (2005)
Genetics 171, 7-21
   Abstract »    Full Text »    PDF »
Nuclear Localization Signal Peptides Induce Molecular Delivery along Microtubules.
H. Salman, A. Abu-Arish, S. Oliel, A. Loyter, J. Klafter, R. Granek, and M. Elbaum (2005)
Biophys. J. 89, 2134-2145
   Abstract »    Full Text »    PDF »
Spatial Coordination of Spindle Assembly by Chromosome-Mediated Signaling Gradients.
M. Caudron, G. Bunt, P. Bastiaens, and E. Karsenti (2005)
Science 309, 1373-1376
   Abstract »    Full Text »    PDF »
Tension-dependent Regulation of Microtubule Dynamics at Kinetochores Can Explain Metaphase Congression in Yeast.
M. K. Gardner, C. G. Pearson, B. L. Sprague, T. R. Zarzar, K. Bloom, E. D. Salmon, and D. J. Odde (2005)
Mol. Biol. Cell 16, 3764-3775
   Abstract »    Full Text »    PDF »
Nuclear Targeting of Adenovirus Type 2 Requires CRM1-mediated Nuclear Export.
S. Strunze, L. C. Trotman, K. Boucke, and U. F. Greber (2005)
Mol. Biol. Cell 16, 2999-3009
   Abstract »    Full Text »    PDF »
Receptor for Hyaluronan-Mediated Motility Correlates with Centrosome Abnormalities in Multiple Myeloma and Maintains Mitotic Integrity.
C. A. Maxwell, J. J. Keats, A. R. Belch, L. M. Pilarski, and T. Reiman (2005)
Cancer Res. 65, 850-860
   Abstract »    Full Text »    PDF »
Importin {beta} is transported to spindle poles during mitosis and regulates Ran-dependent spindle assembly factors in mammalian cells.
M. Ciciarello, R. Mangiacasale, C. Thibier, G. Guarguaglini, E. Marchetti, B. Di Fiore, and P. Lavia (2004)
J. Cell Sci. 117, 6511-6522
   Abstract »    Full Text »    PDF »
Schizosaccharomyces pombe RanGAP Homolog, SpRna1, Is Required for Centromeric Silencing and Chromosome Segregation.
A. Kusano, T. Yoshioka, H. Nishijima, H. Nishitani, and T. Nishimoto (2004)
Mol. Biol. Cell 15, 4960-4970
   Abstract »    Full Text »    PDF »
The mechanism of spindle assembly: functions of Ran and its target TPX2.
O. J. Gruss and I. Vernos (2004)
J. Cell Biol. 166, 949-955
   Abstract »    Full Text »    PDF »
An ATP-dependent Activity That Releases RanGDP from NTF2.
M. Yamada, I. W. Mattaj, and Y. Yoneda (2004)
J. Biol. Chem. 279, 36228-36234
   Abstract »    Full Text »    PDF »
{beta}-Catenin Shows an Overlapping Sequence Requirement but Distinct Molecular Interactions for Its Bidirectional Passage through Nuclear Pores.
M. Koike, S. Kose, M. Furuta, N. Taniguchi, F. Yokoya, Y. Yoneda, and N. Imamoto (2004)
J. Biol. Chem. 279, 34038-34047
   Abstract »    Full Text »    PDF »
Eukaryotic Cells and their Cell Bodies: Cell Theory Revised.
F. BALUSKA, D. VOLKMANN, and P. W. BARLOW (2004)
Ann. Bot. 94, 9-32
   Abstract »    Full Text »    PDF »
Microtubule Plus-End Dynamics in Xenopus Egg Extract Spindles.
J. S. Tirnauer, E. D. Salmon, and T. J. Mitchison (2004)
Mol. Biol. Cell 15, 1776-1784
   Abstract »    Full Text »    PDF »
CRM1 and Ran are present but a NES-CRM1-RanGTP complex is not required in Balbiani ring mRNP particles from the gene to the cytoplasm.
J. Zhao, S.-B. Jin, and L. Wieslander (2004)
J. Cell Sci. 117, 1553-1566
   Abstract »    Full Text »    PDF »
Phosphorylation of RCC1 in mitosis is essential for producing a high RanGTP concentration on chromosomes and for spindle assembly in mammalian cells.
H.-Y. Li and Y. Zheng (2004)
Genes & Dev. 18, 512-527
   Abstract »    Full Text »    PDF »
Importin {alpha}/{beta} and Ran-GTP Regulate XCTK2 Microtubule Binding through a Bipartite Nuclear Localization Signal.
S. C. Ems-McClung, Y. Zheng, and C. E. Walczak (2004)
Mol. Biol. Cell 15, 46-57
   Abstract »    Full Text »    PDF »
Ran modulates spindle assembly by regulating a subset of TPX2 and Kid activities including Aurora A activation.
N. Trieselmann, S. Armstrong, J. Rauw, and A. Wilde (2003)
J. Cell Sci. 116, 4791-4798
   Abstract »    Full Text »    PDF »
Biochemical Characterization of the Ran-RanBP1-RanGAP System: Are RanBP Proteins and the Acidic Tail of RanGAP Required for the Ran-RanGAP GTPase Reaction?.
M. J. Seewald, A. Kraemer, M. Farkasovsky, C. Korner, A. Wittinghofer, and I. R. Vetter (2003)
Mol. Cell. Biol. 23, 8124-8136
   Abstract »    Full Text »    PDF »
Importin {beta} Negatively Regulates Nuclear Membrane Fusion and Nuclear Pore Complex Assembly.
A. Harel, R. C. Chan, A. Lachish-Zalait, E. Zimmerman, M. Elbaum, and D. J. Forbes (2003)
Mol. Biol. Cell 14, 4387-4396
   Abstract »    Full Text »    PDF »
Yeast Kinetochores Do Not Stabilize Stu2p-dependent Spindle Microtubule Dynamics.
C. G. Pearson, P. S. Maddox, T. R. Zarzar, E.D. Salmon, and K. Bloom (2003)
Mol. Biol. Cell 14, 4181-4195
   Abstract »    Full Text »    PDF »
Part of Ran Is Associated with AKAP450 at the Centrosome: Involvement in Microtubule-organizing Activity.
G. Keryer, B. Di Fiore, C. Celati, K. F. Lechtreck, M. Mogensen, A. Delouvee, P. Lavia, M. Bornens, and A.-M. Tassin (2003)
Mol. Biol. Cell 14, 4260-4271
   Abstract »    Full Text »    PDF »
Mammalian RanBP1 regulates centrosome cohesion during mitosis.
B. Di Fiore, M. Ciciarello, R. Mangiacasale, A. Palena, A.-M. Tassin, E. Cundari, and P. Lavia (2003)
J. Cell Sci. 116, 3399-3411
   Abstract »    Full Text »    PDF »
The condensin complex is required for proper spindle assembly and chromosome segregation in Xenopus egg extracts.
S. M. Wignall, R. Deehan, T. J. Maresca, and R. Heald (2003)
J. Cell Biol. 161, 1041-1051
   Abstract »    Full Text »    PDF »
In vitro analysis of nuclear mRNA export using molecular beacons for target detection.
R. H. Kehlenbach (2003)
Nucleic Acids Res. 31, e64
   Abstract »    Full Text »    PDF »
Mechanisms of Microtubule-Based Kinetochore Positioning in the Yeast Metaphase Spindle.
B. L. Sprague, C. G. Pearson, P. S. Maddox, K. S. Bloom, E. D. Salmon, and David. J. Odde (2003)
Biophys. J. 84, 3529-3546
   Abstract »    Full Text »    PDF »
Recombinant Antibodies to the Small GTPase Rab6 as Conformation Sensors.
C. Nizak, S. Monier, E. del Nery, S. Moutel, B. Goud, and F. Perez (2003)
Science 300, 984-987
   Abstract »    Full Text »    PDF »
Controlled Damage in Thick Specimens by Multiphoton Excitation.
J. A. Galbraith and M. Terasaki (2003)
Mol. Biol. Cell 14, 1808-1817
   Abstract »    Full Text »    PDF »
Mitosis Through the Microscope: Advances in Seeing Inside Live Dividing Cells.
C. L. Rieder and A. Khodjakov (2003)
Science 300, 91-96
   Abstract »    Full Text »    PDF »
The Ran GTPase cycle is required for yeast nuclear pore complex assembly.
K. J. Ryan, J. M. McCaffery, and S. R. Wente (2003)
J. Cell Biol. 160, 1041-1053
   Abstract »    Full Text »    PDF »
E1A Deregulates the Centrosome Cycle in a Ran GTPase-dependent Manner.
A. De Luca, R. Mangiacasale, A. Severino, L. Malquori, A. Baldi, A. Palena, A. M. Mileo, P. Lavia, and M. G. Paggi (2003)
Cancer Res. 63, 1430-1437
   Abstract »    Full Text »    PDF »
A mechanism of coupling RCC1 mobility to RanGTP production on the chromatin in vivo.
H. Y. Li, D. Wirtz, and Y. Zheng (2003)
J. Cell Biol. 160, 635-644
   Abstract »    Full Text »    PDF »
The strategy for coupling the RanGTP gradient to nuclear protein export.
A. Becskei and I. W. Mattaj (2003)
PNAS 100, 1717-1722
   Abstract »    Full Text »    PDF »
Shedding Light on Cell Signaling: Interpretation of FRET Biosensors.
F. Gaits and K. Hahn (2003)
Sci. STKE 2003, pe3
   Abstract »    Full Text »    PDF »
Human centromere chromatin protein hMis12, essential for equal segregation, is independent of CENP-A loading pathway.
G. Goshima, T. Kiyomitsu, K. Yoda, and M. Yanagida (2003)
J. Cell Biol. 160, 25-39
   Abstract »    Full Text »    PDF »
The Ran GTPase System in Fission Yeast Affects Microtubules and Cytokinesis in Cells That Are Competent for Nucleocytoplasmic Protein Transport.
S. S. Salus, J. Demeter, and S. Sazer (2002)
Mol. Cell. Biol. 22, 8491-8505
   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 »
Fluorescence Resonance Energy Transfer Biosensors That Detect Ran Conformational Changes and a Ran{middle dot}GDP-Importin-beta -RanBP1 Complex in Vitro and in Intact Cells.
K. Plafker and I. G. Macara (2002)
J. Biol. Chem. 277, 30121-30127
   Abstract »    Full Text »    PDF »
SMN, the spinal muscular atrophy protein, forms a pre-import snRNP complex with snurportin1 and importin {beta}.
U. Narayanan, J. K. Ospina, M. R. Frey, M. D. Hebert, and A. G. Matera (2002)
Hum. Mol. Genet. 11, 1785-1795
   Abstract »    Full Text »    PDF »
Nercc1, a mammalian NIMA-family kinase, binds the Ran GTPase and regulates mitotic progression.
J. Roig, A. Mikhailov, C. Belham, and J. Avruch (2002)
Genes & Dev. 16, 1640-1658
   Abstract »    Full Text »    PDF »
Chromosomal association of Ran during meiotic and mitotic divisions.
B. Hinkle, B. Slepchenko, M. M. Rolls, T. C. Walther, P. A. Stein, L. M. Mehlmann, J. Ellenberg, and M. Terasaki (2002)
J. Cell Sci. 115, 4685-4693
   Abstract »    Full Text »    PDF »


ADVERTISEMENT
Click Me!

ADVERTISEMENT
Click Me!

To Advertise     Find Products

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