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.
Subscribe

Logo for

J. Cell Biol. 148 (5): 849-856

Copyright © 2000 by the Rockefeller University Press.

Brief Report

Nuclear Export of Map Kinase (ERK) Involves a Map Kinase Kinase (Mek-Dependent) Active Transport Mechanism

Makoto Adachia, Makoto Fukudaa,b, , and Eisuke Nishidaa,b

a Department of Biophysics, Graduate School of Science, Graduate School of Biostudies, Kyoto University, Sakyo-ku, Kyoto 606-8502, Japan
b Department of Cell and Developmental Biology, Graduate School of Biostudies, Kyoto University, Sakyo-ku, Kyoto 606-8502, Japan
Department of Biophysics, Graduate School of Science, Kyoto University, Sakyo-ku, Kyoto 606-8502, Japan.81-75-753-423581-75-753-4230

L50174{at}sakura.kudpc.kyoto-u.ac.jp

Abstract: In response to extracellular stimuli, mitogen-activated protein kinase (MAPK, also known as ERK), which localizes to the cytoplasm in quiescent cells, translocates to the nucleus and then relocalizes to the cytoplasm again. The relocalization of nuclear MAPK to the cytoplasm was not inhibited by cycloheximide, confirming that the relocalization is achieved by nuclear export, but not synthesis, of MAPK. The nuclear export of MAPK was inhibited by leptomycin B (LMB), a specific inhibitor of the nuclear export signal (NES)-dependent transport. We have then shown that MAP kinase kinase (MAPKK, also known as MEK), which mostly localizes to the cytoplasm because of its having NES, is able to shuttle between the cytoplasm and the nucleus constantly. MAPK, when injected into the nucleus, was rapidly exported from the nucleus by coinjected wild-type MAPKK, but not by the NES-disrupted MAPKK. In addition, injection of the fragment corresponding to the MAPK-binding site of MAPKK into the nucleus, which would disrupt the binding of MAPK to MAPKK in the nucleus, significantly inhibited the nuclear export of endogenous MAPK. Taken together, these results suggest that the relocalization of nuclear MAPK to the cytoplasm involves a MAPKK-dependent, active transport mechanism.

Key Words: leptomycin B • MAP kinase • nuclear • export • phosphorylation • signal transduction

Abbreviations used in this paper: CHX, cycloheximide; DAPI, 4',6-diamidino-2-phenylindole; LMB, leptomycin B; MAPK, mitogen-activated protein kinase; MAPKK, MAPK kinase; NES, nuclear export signal; TPA, 12-O-tetra decanoyl phorbol myristate acetate.

THIS ARTICLE HAS BEEN CITED BY OTHER ARTICLES:
Sef Downregulation by Ras Causes MEK1/2 to Become Aberrantly Nuclear Localized Leading to Polyploidy and Neoplastic Transformation.
S. Duhamel, J. Hebert, L. Gaboury, A. Bouchard, R. Simon, G. Sauter, M. Basik, and S. Meloche (2012)
Cancer Res. 72, 626-635
   Abstract »    Full Text »    PDF »
Ran Is a Potential Therapeutic Target for Cancer Cells with Molecular Changes Associated with Activation of the PI3K/Akt/mTORC1 and Ras/MEK/ERK Pathways.
H.-F. Yuen, K.-K. Chan, C. Grills, J. T. Murray, A. Platt-Higgins, O. S. Eldin, K. O'Byrne, P. Janne, D. A. Fennell, P. G. Johnston, et al. (2012)
Clin. Cancer Res. 18, 380-391
   Abstract »    Full Text »    PDF »
ERK Nuclear Translocation Is Dimerization-independent but Controlled by the Rate of Phosphorylation.
D. S. Lidke, F. Huang, J. N. Post, B. Rieger, J. Wilsbacher, J. L. Thomas, J. Pouyssegur, T. M. Jovin, and P. Lenormand (2010)
J. Biol. Chem. 285, 3092-3102
   Abstract »    Full Text »    PDF »
Calcium-mediated Interactions Regulate the Subcellular Localization of Extracellular Signal-regulated Kinases.
D. Chuderland, G. Marmor, A. Shainskaya, and R. Seger (2008)
J. Biol. Chem. 283, 11176-11188
   Abstract »    Full Text »    PDF »
MEK1 and MEK2 regulate distinct functions by sorting ERK2 to different intracellular compartments.
E. Skarpen, L. I. Flinder, C. M. Rosseland, S. Orstavik, L. Wierod, M. P. Oksvold, B. S. Skalhegg, and H. S. Huitfeldt (2008)
FASEB J 22, 466-476
   Abstract »    Full Text »    PDF »
Single and Combined Silencing of ERK1 and ERK2 Reveals Their Positive Contribution to Growth Signaling Depending on Their Expression Levels.
R. Lefloch, J. Pouyssegur, and P. Lenormand (2008)
Mol. Cell. Biol. 28, 511-527
   Abstract »    Full Text »    PDF »
Mutations in ERK2 Binding Sites Affect Nuclear Entry.
M. N. Yazicioglu, D. L. Goad, A. Ranganathan, A. W. Whitehurst, E. J. Goldsmith, and M. H. Cobb (2007)
J. Biol. Chem. 282, 28759-28767
   Abstract »    Full Text »    PDF »
Tamoxifen-Induced Rapid Death of MCF-7 Breast Cancer Cells Is Mediated via Extracellularly Signal-Regulated Kinase Signaling and Can Be Abrogated by Estrogen.
A. Zheng, A. Kallio, and P. Harkonen (2007)
Endocrinology 148, 2764-2777
   Abstract »    Full Text »    PDF »
Activation and nuclear translocation of ERK in response to ligand-dependent and -independent stimuli in liver and gill cells from rainbow trout.
H. L. Ebner, M. Blatzer, M. Nawaz, and G. Krumschnabel (2007)
J. Exp. Biol. 210, 1036-1045
   Abstract »    Full Text »    PDF »
Interaction with MEK Causes Nuclear Export and Downregulation of Peroxisome Proliferator-Activated Receptor {gamma}.
E. Burgermeister, D. Chuderland, T. Hanoch, M. Meyer, M. Liscovitch, and R. Seger (2007)
Mol. Cell. Biol. 27, 803-817
   Abstract »    Full Text »    PDF »
Dynamic regulation of ERK2 nuclear translocation and mobility in living cells.
M. Costa, M. Marchi, F. Cardarelli, A. Roy, F. Beltram, L. Maffei, and G. M. Ratto (2006)
J. Cell Sci. 119, 4952-4963
   Abstract »    Full Text »    PDF »
The Nuclear Localization of ERK2 Occurs by Mechanisms Both Independent of and Dependent on Energy.
A. Ranganathan, M. N. Yazicioglu, and M. H. Cobb (2006)
J. Biol. Chem. 281, 15645-15652
   Abstract »    Full Text »    PDF »
Dynamics of the Ras/ERK MAPK Cascade as Monitored by Fluorescent Probes.
A. Fujioka, K. Terai, R. E. Itoh, K. Aoki, T. Nakamura, S. Kuroda, E. Nishida, and M. Matsuda (2006)
J. Biol. Chem. 281, 8917-8926
   Abstract »    Full Text »    PDF »
Arrestin-mediated ERK Activation by Gonadotropin-releasing Hormone Receptors: RECEPTOR-SPECIFIC ACTIVATION MECHANISMS AND COMPARTMENTALIZATION.
C. J. Caunt, A. R. Finch, K. R. Sedgley, L. Oakley, L. M. Luttrell, and C. A. McArdle (2006)
J. Biol. Chem. 281, 2701-2710
   Abstract »    Full Text »    PDF »
Live Cell Imaging of ERK and MEK: SIMPLE BINDING EQUILIBRIUM EXPLAINS THE REGULATED NUCLEOCYTOPLASMIC DISTRIBUTION OF ERK.
W. R. Burack and A. S. Shaw (2005)
J. Biol. Chem. 280, 3832-3837
   Abstract »    Full Text »    PDF »
Stimulus-Coupled Spatial Restriction of Extracellular Signal-Regulated Kinase 1/2 Activity Contributes to the Specificity of Signal-Response Pathways.
A. Whitehurst, M. H. Cobb, and M. A. White (2004)
Mol. Cell. Biol. 24, 10145-10150
   Abstract »    Full Text »    PDF »
Regulatory Mechanisms and Function of ERK MAP Kinases.
S. Torii, K. Nakayama, T. Yamamoto, and E. Nishida (2004)
J. Biochem. 136, 557-561
   Abstract »    Full Text »    PDF »
Induction of ERK1/2 and Histone H3 Phosphorylation within the Outer Stripe of the Outer Medulla of the Eker Rat by 2,3,5-Tris-(Glutathion-S-yl)hydroquinone.
J. Dong, J. I. Everitt, S. S. Lau, and T. J. Monks (2004)
Toxicol. Sci. 80, 350-357
   Abstract »    Full Text »    PDF »
ERK1/2 Associates with the c-Met-binding Domain of Growth Factor Receptor-bound Protein 2 (Grb2)-associated Binder-1 (Gab1): ROLE IN ERK1/2 AND EARLY GROWTH RESPONSE FACTOR-1 (Egr-1) NUCLEAR ACCUMULATION.
M. Osawa, S. Itoh, S. Ohta, Q. Huang, B. C. Berk, N.-L. Marmarosh, W. Che, B. Ding, C. Yan, and J.-i. Abe (2004)
J. Biol. Chem. 279, 29691-29699
   Abstract »    Full Text »    PDF »
Regulation of Ras-MAPK pathway mitogenic activity by restricting nuclear entry of activated MAPK in endoderm differentiation of embryonic carcinoma and stem cells.
E. R. Smith, J. L. Smedberg, M. E. Rula, and X.-X. Xu (2004)
J. Cell Biol. 164, 689-699
   Abstract »    Full Text »    PDF »
ERF Nuclear Shuttling, a Continuous Monitor of Erk Activity That Links It to Cell Cycle Progression.
L. Le Gallic, L. Virgilio, P. Cohen, B. Biteau, and G. Mavrothalassitis (2004)
Mol. Cell. Biol. 24, 1206-1218
   Abstract »    Full Text »    PDF »
Altered Mitogen-Activated Protein Kinase Signal Transduction in Human Skin Fibroblasts During In Vitro Aging: Differential Expression of Low-Density Lipoprotein Receptor.
C. Bose, C. Bhuvaneswaran, and K. B. Udupa (2004)
J Gerontol A Biol Sci Med Sci 59, B126-B135
   Abstract »    Full Text »    PDF »
Nuclear Export of ERK3 by a CRM1-dependent Mechanism Regulates Its Inhibitory Action on Cell Cycle Progression.
C. Julien, P. Coulombe, and S. Meloche (2003)
J. Biol. Chem. 278, 42615-42624
   Abstract »    Full Text »    PDF »
CRM1/Ran-Mediated Nuclear Export of p27Kip1 Involves a Nuclear Export Signal and Links p27 Export and Proteolysis.
M. K. Connor, R. Kotchetkov, S. Cariou, A. Resch, R. Lupetti, R. G. Beniston, F. Melchior, L. Hengst, and J. M. Slingerland (2003)
Mol. Biol. Cell 14, 201-213
   Abstract »    Full Text »
The NS2 Proteins of Parvovirus Minute Virus of Mice Are Required for Efficient Nuclear Egress of Progeny Virions in Mouse Cells.
V. Eichwald, L. Daeffler, M. Klein, J. Rommelaere, and N. Salome (2002)
J. Virol. 76, 10307-10319
   Abstract »    Full Text »    PDF »
Nuclear import of factors involved in signaling is inhibited in C3H/10T1/2 cells treated with tetradecylthioacetic acid.
B. Bjorndal, C. Helleland, S.-O. Boe, O. A. Gudbrandsen, K.-H. Kalland, P. Bohov, R. K. Berge, and J. R. Lillehaug (2002)
J. Lipid Res. 43, 1630-1640
   Abstract »    Full Text »    PDF »
Identification of a C-terminal Region That Is Required for the Nuclear Translocation of ERK2 by Passive Diffusion.
S. Shibayama, R. Shibata-Seita, K. Miura, Y. Kirino, and K. Takishima (2002)
J. Biol. Chem. 277, 37777-37782
   Abstract »    Full Text »    PDF »
MEK1 Signaling Mediates Transformation and Metastasis of EpH4 Mammary Epithelial Cells Independent of an Epithelial to Mesenchymal Transition.
J. Pinkas and P. Leder (2002)
Cancer Res. 62, 4781-4790
   Abstract »    Full Text »    PDF »
Cytoplasmic Localization of Wis1 MAPKK by Nuclear Export Signal Is Important for Nuclear Targeting of Spc1/Sty1 MAPK in Fission Yeast.
A. N. Nguyen, A. D. Ikner, M. Shiozaki, S. M. Warren, and K. Shiozaki (2002)
Mol. Biol. Cell 13, 2651-2663
   Abstract »    Full Text »    PDF »
Epidermal Growth Factor-mediated Activation of the ETS Domain Transcription Factor Elk-1 Requires Nuclear Calcium.
T. Pusl, J. J. Wu, T. L. Zimmerman, L. Zhang, B. E. Ehrlich, M. W. Berchtold, J. B. Hoek, S. J. Karpen, M. H. Nathanson, and A. M. Bennett (2002)
J. Biol. Chem. 277, 27517-27527
   Abstract »    Full Text »    PDF »
ERK2 enters the nucleus by a carrier-independent mechanism.
A. W. Whitehurst, J. L. Wilsbacher, Y. You, K. Luby-Phelps, M. S. Moore, and M. H. Cobb (2002)
PNAS 99, 7496-7501
   Abstract »    Full Text »    PDF »
AT1 Receptor Mutant Lacking Heterotrimeric G Protein Coupling Activates the Src-Ras-ERK Pathway without Nuclear Translocation of ERKs.
K. Seta, M. Nanamori, J. G. Modrall, R. R. Neubig, and J. Sadoshima (2002)
J. Biol. Chem. 277, 9268-9277
   Abstract »    Full Text »    PDF »
Selective in Vivo Inhibition of Mitogen-activated Protein Kinase Activation Using Cell-permeable Peptides.
B. R. Kelemen, K. Hsiao, and S. A. Goueli (2002)
J. Biol. Chem. 277, 8741-8748
   Abstract »    Full Text »    PDF »
Different Domains of the Mitogen-activated Protein Kinases ERK3 and ERK2 Direct Subcellular Localization and Upstream Specificity in Vivo.
M. J. Robinson, B.-e Xu, S. Stippec, and M. H. Cobb (2002)
J. Biol. Chem. 277, 5094-5100
   Abstract »    Full Text »    PDF »
Phosphorylation Regulates the Nucleocytoplasmic Distribution of Kinase Suppressor of Ras.
J. A. Brennan, D. J. Volle, O. V. Chaika, and R. E. Lewis (2002)
J. Biol. Chem. 277, 5369-5377
   Abstract »    Full Text »    PDF »
Prolonged Nuclear Retention of Activated Extracellular Signal-regulated Protein Kinase Promotes Cell Death Generated by Oxidative Toxicity or Proteasome Inhibition in a Neuronal Cell Line.
M. Stanciu and D. B. DeFranco (2002)
J. Biol. Chem. 277, 4010-4017
   Abstract »    Full Text »    PDF »
Visna Virus-Induced Activation of MAPK Is Required for Virus Replication and Correlates with Virus-Induced Neuropathology.
S. A. Barber, L. Bruett, B. R. Douglass, D. S. Herbst, M. C. Zink, and J. E. Clements (2002)
J. Virol. 76, 817-828
   Abstract »    Full Text »    PDF »
Evidence for Existence of a Nuclear Pore Complex-mediated, Cytosol-independent Pathway of Nuclear Translocation of ERK MAP Kinase in Permeabilized Cells.
Y. Matsubayashi, M. Fukuda, and E. Nishida (2001)
J. Biol. Chem. 276, 41755-41760
   Abstract »    Full Text »    PDF »
MKP-7, a Novel Mitogen-activated Protein Kinase Phosphatase, Functions as a Shuttle Protein.
K. Masuda, H. Shima, M. Watanabe, and K. Kikuchi (2001)
J. Biol. Chem. 276, 39002-39011
   Abstract »    Full Text »    PDF »
The nucleus, a site for signal termination by sequestration and inactivation of p42/p44 MAP kinases.
V. Volmat, M. Camps, S. Arkinstall, J. Pouyssegur, and P. Lenormand (2001)
J. Cell Sci. 114, 3433-3443
   Abstract »    Full Text »    PDF »
Integrin-Mediated Adhesion Regulates ERK Nuclear Translocation and Phosphorylation of Elk-1.
A. E. Aplin, S. A. Stewart, R. K. Assoian, and R.L. Juliano (2001)
J. Cell Biol. 153, 273-282
   Abstract »    Full Text »    PDF »
Mitogen-Activated Protein (MAP) Kinase Pathways: Regulation and Physiological Functions.
G. Pearson, F. Robinson, T. Beers Gibson, B.-e Xu, M. Karandikar, K. Berman, and M. H. Cobb (2001)
Endocr. Rev. 22, 153-183
   Abstract »    Full Text »    PDF »
Beclin 1 Contains a Leucine-rich Nuclear Export Signal That Is Required for Its Autophagy and Tumor Suppressor Function.
X. H. Liang, J. Yu, K. Brown, and B. Levine (2001)
Cancer Res. 61, 3443-3449
   Abstract »    Full Text »
MST, a Physiological Caspase Substrate, Highly Sensitizes Apoptosis Both Upstream and Downstream of Caspase Activation.
K.-K. Lee, T. Ohyama, N. Yajima, S. Tsubuki, and S. Yonehara (2001)
J. Biol. Chem. 276, 19276-19285
   Abstract »    Full Text »    PDF »
Regulation of Ribosomal S6 Kinase 2 by Effectors of the Phosphoinositide 3-Kinase Pathway.
K. A. Martin, S. S. Schalm, C. Richardson, A. Romanelli, K. L. Keon, and J. Blenis (2001)
J. Biol. Chem. 276, 7884-7891
   Abstract »    Full Text »    PDF »
Phosphoinositide 3-Kinase-dependent Regulation of Interleukin-3-induced Proliferation. INVOLVEMENT OF MITOGEN-ACTIVATED PROTEIN KINASES, SHP2 AND Gab2.
B. L. Craddock, J. Hobbs, C. E. Edmead, and M. J. Welham (2001)
J. Biol. Chem. 276, 24274-24283
   Abstract »    Full Text »    PDF »
A Conserved Docking Site in MEKs Mediates High-affinity Binding to MAP Kinases and Cooperates with a Scaffold Protein to Enhance Signal Transmission.
A. J. Bardwell, L. J. Flatauer, K. Matsukuma, J. Thorner, and L. Bardwell (2001)
J. Biol. Chem. 276, 10374-10386
   Abstract »    Full Text »    PDF »
Involvement of the Activation Loop of ERK in the Detachment from Cytosolic Anchoring.
I. Wolf, H. Rubinfeld, S. Yoon, G. Marmor, T. Hanoch, and R. Seger (2001)
J. Biol. Chem. 276, 24490-24497
   Abstract »    Full Text »    PDF »
Regulation of Nuclear Localization during Signaling.
M. S. Cyert (2001)
J. Biol. Chem. 276, 20805-20808
   Full Text »    PDF »

To Advertise     Find Products

Science Signaling. ISSN 1937-9145 (online), 1945-0877 (print). Pre-2008: Science's STKE. ISSN 1525-8882