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 30 November 2001:
Vol. 294. no. 5548, pp. 1895 - 1901
DOI: 10.1126/science.1064693
Prev | Table of Contents | Next

Research Articles

Delineation of mRNA Export Pathways by the Use of Cell-Permeable Peptides

Imed-Eddine Gallouzi,* Joan A. Steitzdagger

The transport of messenger RNAs (mRNAs) from the nucleus to the cytoplasm involves adapter proteins that bind the mRNA as well as receptor proteins that interact with the nuclear pore complex. We demonstrate the utility of cell-permeable peptides designed to interfere with interactions between potential adapter and receptor proteins to define the pathways accessed by particular mRNAs. We show that HuR, a protein implicated in the stabilization of short-lived mRNAs containing AU-rich elements (AREs), serves as an adapter for c-fos mRNA export through two pathways. One involves the HuR shuttling domain, HNS, which exhibits a heat shock-sensitive interaction with transportin 2 (Trn2); the other involves two protein ligands of HuR--pp32 and APRIL--which contain leucine-rich nuclear export signals (NES) recognized by the export receptor CRM1. Heterokaryon and in situ hybridization experiments reveal that the peptides selectively block the nucleocytoplasmic shuttling of their respective adapter proteins without perturbing the overall cellular distribution of polyadenylated mRNAs.

Department of Molecular Biophysics and Biochemistry, Howard Hughes Medical Institute, Yale University School of Medicine, 295 Congress Avenue, New Haven, CT 06536, USA.
*   Present address: Department of Biochemistry, McGill University, McIntyre Medical Sciences Building, 3655 Promenade Sir William Osler, Montreal, Quebec H3G 1Y6, Canada.

dagger    To whom correspondence should be addressed: E-mail: joan.steitz{at}yale.edu

Read the Full Text


THIS ARTICLE HAS BEEN CITED BY OTHER ARTICLES:
Nuclear HuR accumulation through phosphorylation by Cdk1.
H. H. Kim, K. Abdelmohsen, A. Lal, R. Pullmann Jr., X. Yang, S. Galban, S. Srikantan, J. L. Martindale, J. Blethrow, K. M. Shokat, et al. (2008)
Genes & Dev. 22, 1804-1815
   Abstract »    Full Text »    PDF »
Caspase-mediated cleavage of HuR in the cytoplasm contributes to pp32/PHAP-I regulation of apoptosis.
R. Mazroui, S. Di Marco, E. Clair, C. von Roretz, S. A. Tenenbaum, J. D. Keene, M. Saleh, and I.-E. Gallouzi (2008)
J. Cell Biol. 180, 113-127
   Abstract »    Full Text »    PDF »
Prognostic Role of HuR in Hereditary Breast Cancer.
M. Heinonen, R. Fagerholm, K. Aaltonen, O. Kilpivaara, K. Aittomaki, C. Blomqvist, P. Heikkila, C. Haglund, H. Nevanlinna, and A. Ristimaki (2007)
Clin. Cancer Res. 13, 6959-6963
   Abstract »    Full Text »    PDF »
The RNA-Binding Protein HuR Promotes Cell Migration and Cell Invasion by Stabilizing the {beta}-actin mRNA in a U-Rich-Element-Dependent Manner.
V. Dormoy-Raclet, I. Menard, E. Clair, G. Kurban, R. Mazroui, S. Di Marco, C. von Roretz, A. Pause, and I.-E. Gallouzi (2007)
Mol. Cell. Biol. 27, 5365-5380
   Abstract »    Full Text »    PDF »
Specific Protein Domains Mediate Cooperative Assembly of HuR Oligomers on AU-rich mRNA-destabilizing Sequences.
E. J. Fialcowitz-White, B. Y. Brewer, J. D. Ballin, C. D. Willis, E. A. Toth, and G. M. Wilson (2007)
J. Biol. Chem. 282, 20948-20959
   Abstract »    Full Text »    PDF »
Involvement of Transportin 2-mediated HuR Import in Muscle Cell Differentiation.
K. van der Giessen and I.-E. Gallouzi (2007)
Mol. Biol. Cell 18, 2619-2629
   Abstract »    Full Text »    PDF »
Enhanced Replication of Human T-Cell Leukemia Virus Type 1 in T Cells from Transgenic Rats Expressing Human CRM1 That Is Regulated in a Natural Manner.
R. Takayanagi, T. Ohashi, E. Yamashita, Y. Kurosaki, K. Tanaka, Y. Hakata, Y. Komoda, S. Ikeda, Y. Tsunetsugu-Yokota, Y. Tanaka, et al. (2007)
J. Virol. 81, 5908-5918
   Abstract »    Full Text »    PDF »
Analysis of Nucleocytoplasmic Trafficking of the HuR Ligand APRIL and Its Influence on CD83 Expression.
B. Fries, J. Heukeshoven, I. Hauber, C. Gruttner, C. Stocking, R. H. Kehlenbach, J. Hauber, and J. Chemnitz (2007)
J. Biol. Chem. 282, 4504-4515
   Abstract »    Full Text »    PDF »
Differential IFN-{gamma} Stimulation of HLA-A Gene Expression through CRM-1-Dependent Nuclear RNA Export.
S. K. Browne, J. R. Roesser, S. Z. Zhu, and G. D. Ginder (2006)
J. Immunol. 177, 8612-8619
   Abstract »    Full Text »    PDF »
HuD Distribution Changes in Response to Heat Shock but Not Neurotrophic Stimulation.
R. W. Burry and C. L. Smith (2006)
J. Histochem. Cytochem. 54, 1129-1138
   Abstract »    Full Text »    PDF »
Expression of CD83 Is Regulated by HuR via a Novel cis-Active Coding Region RNA Element.
A. T. Prechtel, J. Chemnitz, S. Schirmer, C. Ehlers, I. Langbein-Detsch, J. Stulke, M.-C. Dabauvalle, R. H. Kehlenbach, and J. Hauber (2006)
J. Biol. Chem. 281, 10912-10925
   Abstract »    Full Text »    PDF »
LFA-1-Dependent HuR Nuclear Export and Cytokine mRNA Stabilization in T Cell Activation.
J. G. Wang, M. Collinge, V. Ramgolam, O. Ayalon, X. C. Fan, R. Pardi, and J. R. Bender (2006)
J. Immunol. 176, 2105-2113
   Abstract »    Full Text »    PDF »
AU-rich elements and associated factors: are there unifying principles?.
C. Barreau, L. Paillard, and H. B. Osborne (2006)
Nucleic Acids Res. 33, 7138-7150
   Abstract »    Full Text »    PDF »
Identification of the sequence determinants mediating the nucleo-cytoplasmic shuttling of TIAR and TIA-1 RNA-binding proteins.
T. Zhang, N. Delestienne, G. Huez, V. Kruys, and C. Gueydan (2005)
J. Cell Sci. 118, 5453-5463
   Abstract »    Full Text »    PDF »
p38 Mitogen-Activated Protein Kinase/Hog1p Regulates Translation of the AU-Rich-Element-Bearing MFA2 Transcript.
S. Vasudevan, N. Garneau, D. Tu Khounh, and S. W. Peltz (2005)
Mol. Cell. Biol. 25, 9753-9763
   Abstract »    Full Text »    PDF »
Shared RNA-binding sites for interacting members of the Drosophila ELAV family of neuronal proteins.
C. D. Borgeson and M.-L. Samson (2005)
Nucleic Acids Res. 33, 6372-6383
   Abstract »    Full Text »    PDF »
Cyclooxygenase-2 Is an Independent Prognostic Factor in Gastric Cancer and Its Expression Is Regulated by the Messenger RNA Stability Factor HuR.
J. Mrena, J.-P. Wiksten, A. Thiel, A. Kokkola, L. Pohjola, J. Lundin, S. Nordling, A. Ristimaki, and C. Haglund (2005)
Clin. Cancer Res. 11, 7362-7368
   Abstract »    Full Text »    PDF »
RNA-Binding Protein HuR Is Required for Stabilization of SLC11A1 mRNA and SLC11A1 Protein Expression.
Y. Z. Xu, S. Di Marco, I. Gallouzi, M. Rola-Pleszczynski, and D. Radzioch (2005)
Mol. Cell. Biol. 25, 8139-8149
   Abstract »    Full Text »    PDF »
Tudor Domains Bind Symmetrical Dimethylated Arginines.
J. Cote and S. Richard (2005)
J. Biol. Chem. 280, 28476-28483
   Abstract »    Full Text »    PDF »
NF-{kappa}B-Mediated MyoD Decay during Muscle Wasting Requires Nitric Oxide Synthase mRNA Stabilization, HuR Protein, and Nitric Oxide Release.
S. Di Marco, R. Mazroui, P. Dallaire, S. Chittur, S. A. Tenenbaum, D. Radzioch, A. Marette, and I.-E. Gallouzi (2005)
Mol. Cell. Biol. 25, 6533-6545
   Abstract »    Full Text »    PDF »
Adenovirus E4orf6 targets pp32/LANP to control the fate of ARE-containing mRNAs by perturbing the CRM1-dependent mechanism.
F. Higashino, M. Aoyagi, A. Takahashi, M. Ishino, M. Taoka, T. Isobe, M. Kobayashi, Y. Totsuka, T. Kohgo, and M. Shindoh (2005)
J. Cell Biol. 170, 15-20
   Abstract »    Full Text »    PDF »
An Early Event in Adipogenesis, the Nuclear Selection of the CCAAT Enhancer-binding Protein {beta} (C/EBP{beta}) mRNA by HuR and its Translocation to the Cytosol.
K. Gantt, J. Cherry, R. Tenney, V. Karschner, and P. H. Pekala (2005)
J. Biol. Chem. 280, 24768-24774
   Abstract »    Full Text »    PDF »
MEK-ERK Signaling Controls Hdm2 Oncoprotein Expression by Regulating hdm2 mRNA Export to the Cytoplasm.
M. Phelps, A. Phillips, M. Darley, and J. P. Blaydes (2005)
J. Biol. Chem. 280, 16651-16658
   Abstract »    Full Text »    PDF »
tRNA-linked molecular beacons for imaging mRNAs in the cytoplasm of living cells.
M. M. Mhlanga, D. Y. Vargas, C. W. Fung, F. R. Kramer, and S. Tyagi (2005)
Nucleic Acids Res. 33, 1902-1912
   Abstract »    Full Text »    PDF »
Fusion Polypeptides That Inhibit Exocytosis: Fusing Aptamer and Cell-Penetrating Peptide Technologies and Pharmacologies.
L. E. Eiden (2005)
Mol. Pharmacol. 67, 980-982
   Abstract »    Full Text »    PDF »
Cytoplasmic HuR Expression Is a Prognostic Factor in Invasive Ductal Breast Carcinoma.
M. Heinonen, P. Bono, K. Narko, S.-H. Chang, J. Lundin, H. Joensuu, H. Furneaux, T. Hla, C. Haglund, and A. Ristimaki (2005)
Cancer Res. 65, 2157-2161
   Abstract »    Full Text »    PDF »
ACE gene titration in mice uncovers a new mechanism for ACE on the control of body weight.
A. S. Heimann, M. H. Favarato, F. C. Gozzo, V. Rioli, F. R. Carreno, M. N. Eberlin, E. S. Ferro, J. H. Krege, and J. E. Krieger (2005)
Physiol Genomics 20, 173-182
   Abstract »    Full Text »    PDF »
AMP-activated Protein Kinase-regulated Phosphorylation and Acetylation of Importin {alpha}1: INVOLVEMENT IN THE NUCLEAR IMPORT OF RNA-BINDING PROTEIN HuR.
W. Wang, X. Yang, T. Kawai, I. L. de Silanes, K. Mazan-Mamczarz, P. Chen, Y. M. Chook, C. Quensel, M. Kohler, and M. Gorospe (2004)
J. Biol. Chem. 279, 48376-48388
   Abstract »    Full Text »    PDF »
ATP-binding Cassette Transporter A1 Contains a Novel C-terminal VFVNFA Motif That Is Required for Its Cholesterol Efflux and ApoA-I Binding Activities.
M. L. Fitzgerald, K.-i. Okuhira, G. F. Short III, J. J. Manning, S. A. Bell, and M. W. Freeman (2004)
J. Biol. Chem. 279, 48477-48485
   Abstract »    Full Text »    PDF »
Distinct Domains of AU-Rich Elements Exert Different Functions in mRNA Destabilization and Stabilization by p38 Mitogen-Activated Protein Kinase or HuR.
R. Winzen, G. Gowrishankar, F. Bollig, N. Redich, K. Resch, and H. Holtmann (2004)
Mol. Cell. Biol. 24, 4835-4847
   Abstract »    Full Text »    PDF »
CRM1-dependent, but not ARE-mediated, nuclear export of IFN-{alpha}1 mRNA.
T. Kimura, I. Hashimoto, T. Nagase, and J.-I. Fujisawa (2004)
J. Cell Sci. 117, 2259-2270
   Abstract »    Full Text »    PDF »
Generation and Characterization of LANP/pp32 Null Mice.
P. Opal, J. J. Garcia, A. E. McCall, B. Xu, E. J. Weeber, J. D. Sweatt, H. T. Orr, and H. Y. Zoghbi (2004)
Mol. Cell. Biol. 24, 3140-3149
   Abstract »    Full Text »    PDF »
The C-Terminal Transmembrane Domain of Hepatitis C Virus (HCV) RNA Polymerase Is Essential for HCV Replication In Vivo.
K. J. Lee, J. Choi, J.-h. Ou, and M. M. C. Lai (2004)
J. Virol. 78, 3797-3802
   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 »
From The Cover: Transportin2 functions as importin and mediates nuclear import of HuR.
S. Guttinger, P. Muhlhausser, R. Koller-Eichhorn, J. Brennecke, and U. Kutay (2004)
PNAS 101, 2918-2923
   Abstract »    Full Text »    PDF »
Identification of a target RNA motif for RNA-binding protein HuR.
I. L. de Silanes, M. Zhan, A. Lal, X. Yang, and M. Gorospe (2004)
PNAS 101, 2987-2992
   Abstract »    Full Text »    PDF »
The mRNA Export Factor Human Gle1 Interacts with the Nuclear Pore Complex Protein Nup155.
H. J. Rayala, F. Kendirgi, D. M. Barry, P. W. Majerus, and S. R. Wente (2004)
Mol. Cell. Proteomics 3, 145-155
   Abstract »    Full Text »    PDF »
hRIP, a cellular cofactor for Rev function, promotes release of HIV RNAs from the perinuclear region.
N. Sanchez-Velar, E. B. Udofia, Z. Yu, and M. L. Zapp (2004)
Genes & Dev. 18, 23-34
   Abstract »    Full Text »    PDF »
A Multifunctional Domain in Human CRM1 (Exportin 1) Mediates RanBP3 Binding and Multimerization of Human T-Cell Leukemia Virus Type 1 Rex Protein.
Y. Hakata, M. Yamada, and H. Shida (2003)
Mol. Cell. Biol. 23, 8751-8761
   Abstract »    Full Text »    PDF »
RNAi-mediated HuR Depletion Leads to the Inhibition of Muscle Cell Differentiation.
K. van der Giessen, S. Di-Marco, E. Clair, and I. E. Gallouzi (2003)
J. Biol. Chem. 278, 47119-47128
   Abstract »    Full Text »    PDF »
Cytoplasmic HuR Expression Correlates with Poor Outcome and with Cyclooxygenase 2 Expression in Serous Ovarian Carcinoma.
T.-L. Erkinheimo, H. Lassus, A. Sivula, S. Sengupta, H. Furneaux, T. Hla, C. Haglund, R. Butzow, and A. Ristimaki (2003)
Cancer Res. 63, 7591-7594
   Abstract »    Full Text »    PDF »
Structure-based Incorporation of 6-Methyl-8-(2-deoxy-{beta}-ribofuranosyl)isoxanthopteridine into the Human Telomeric Repeat DNA as a Probe for UP1 Binding and Destabilization of G-tetrad Structures.
J. C. Myers, S. A. Moore, and Y. Shamoo (2003)
J. Biol. Chem. 278, 42300-42306
   Abstract »    Full Text »    PDF »
A Novel Nuclear Localization Signal in the Auxiliary Domain of Apobec-1 Complementation Factor Regulates Nucleocytoplasmic Import and Shuttling.
V. Blanc, S. Kennedy, and N. O. Davidson (2003)
J. Biol. Chem. 278, 41198-41204
   Abstract »    Full Text »    PDF »
Nuclear Pnn/DRS Protein Binds to Spliced mRNPs and Participates in mRNA Processing and Export via Interaction with RNPS1.
C. Li, R.-I. Lin, M.-C. Lai, P. Ouyang, and W.-Y. Tarn (2003)
Mol. Cell. Biol. 23, 7363-7376
   Abstract »    Full Text »    PDF »
ELAV inhibits 3'-end processing to promote neural splicing of ewg pre-mRNA.
M. Soller and K. White (2003)
Genes & Dev. 17, 2526-2538
   Abstract »    Full Text »    PDF »
Chronic Ethanol Exposure Increases the Binding of HuR to the TNF{alpha} 3'-Untranslated Region in Macrophages.
M. R. McMullen, E. Cocuzzi, M. Hatzoglou, and L. E. Nagy (2003)
J. Biol. Chem. 278, 38333-38341
   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 »
Gene Expression Profiles Obtained from Fine-Needle Aspirations of Breast Cancer Reliably Identify Routine Prognostic Markers and Reveal Large-Scale Molecular Differences between Estrogen-negative and Estrogen-positive Tumors.
L. Pusztai, M. Ayers, J. Stec, E. Clark, K. Hess, D. Stivers, A. Damokosh, N. Sneige, T. A. Buchholz, F. J. Esteva, et al. (2003)
Clin. Cancer Res. 9, 2406-2415
   Abstract »    Full Text »    PDF »
The RNA-binding Protein HuR Regulates the Expression of Cyclooxygenase-2.
S. Sengupta, B.-C. Jang, M.-T. Wu, J.-H. Paik, H. Furneaux, and T. Hla (2003)
J. Biol. Chem. 278, 25227-25233
   Abstract »    Full Text »    PDF »
Complex Formation among the RNA Export Proteins Nup98, Rae1/Gle2, and TAP.
M. B. Blevins, A. M. Smith, E. M. Phillips, and M. A. Powers (2003)
J. Biol. Chem. 278, 20979-20988
   Abstract »    Full Text »    PDF »
A Constitutive Decay Element Promotes Tumor Necrosis Factor Alpha mRNA Degradation via an AU-Rich Element-Independent Pathway.
G. Stoecklin, M. Lu, B. Rattenbacher, and C. Moroni (2003)
Mol. Cell. Biol. 23, 3506-3515
   Abstract »    Full Text »    PDF »
A quantitative analysis of intron effects on mammalian gene expression.
A. NOTT, S. H. MEISLIN, and M. J. MOORE (2003)
RNA 9, 607-617
   Abstract »    Full Text »    PDF »
An essential role for hGle1 nucleocytoplasmic shuttling in mRNA export.
F. Kendirgi, D. M. Barry, E. R. Griffis, M. A. Powers, and S. R. Wente (2003)
J. Cell Biol. 160, 1029-1040
   Abstract »    Full Text »    PDF »
Protein composition of human prespliceosomes isolated by a tobramycin affinity-selection method.
K. Hartmuth, H. Urlaub, H.-P. Vornlocher, C. L. Will, M. Gentzel, M. Wilm, and R. Luhrmann (2002)
PNAS 99, 16719-16724
   Abstract »    Full Text »    PDF »
ELAV/Hu proteins inhibit p27 translation via an IRES element in the p27 5'UTR.
M. Kullmann, U. Gopfert, B. Siewe, and L. Hengst (2002)
Genes & Dev. 16, 3087-3099
   Abstract »    Full Text »    PDF »
Lipopolysaccharide-induced Methylation of HuR, an mRNA-stabilizing Protein, by CARM1.
H. Li, S. Park, B. Kilburn, M. A. Jelinek, A. Henschen-Edman, D. W. Aswad, M. R. Stallcup, and I. A. Laird-Offringa (2002)
J. Biol. Chem. 277, 44623-44630
   Abstract »    Full Text »    PDF »
Karyopherin beta 2B participates in mRNA export from the nucleus.
M. K. Shamsher, J. Ploski, and A. Radu (2002)
PNAS 99, 14195-14199
   Abstract »    Full Text »    PDF »
Inhibition of Translation by UAUUUAU and UAUUUUUAU Motifs of the AU-rich RNA Instability Element in the HPV-1 Late 3' Untranslated Region.
L. Wiklund, M. Sokolowski, A. Carlsson, M. Rush, and S. Schwartz (2002)
J. Biol. Chem. 277, 40462-40471
   Abstract »    Full Text »    PDF »
Analysis of the Adenovirus E1B-55K-Anchored Proteome Reveals Its Link to Ubiquitination Machinery.
J. N. Harada, A. Shevchenko, A. Shevchenko, D. C. Pallas, and A. J. Berk (2002)
J. Virol. 76, 9194-9206
   Abstract »    Full Text »    PDF »
Novel Binding of HuR and Poly(C)-binding Protein to a Conserved UC-rich Motif within the 3'-Untranslated Region of the Androgen Receptor Messenger RNA.
B. B. Yeap, D. C. Voon, J. P. Vivian, R. K. McCulloch, A. M. Thomson, K. M. Giles, M. F. Czyzyk-Krzeska, H. Furneaux, M. C. J. Wilce, J. A. Wilce, et al. (2002)
J. Biol. Chem. 277, 27183-27192
   Abstract »    Full Text »    PDF »
A fragment of the HMGN2 protein homes to the nuclei of tumor cells and tumor endothelial cells in vivo.
K. Porkka, P. Laakkonen, J. A. Hoffman, M. Bernasconi, and E. Ruoslahti (2002)
PNAS 99, 7444-7449
   Abstract »    Full Text »    PDF »
A Los1p-independent pathway for nuclear export of intronless tRNAs in Saccharomycescerevisiae.
W. Feng and A. K. Hopper (2002)
PNAS 99, 5412-5417
   Abstract »    Full Text »    PDF »
CRM1-Dependent Function of a cis-Acting RNA Export Element.
I. Popa, M. E. Harris, J. E. Donello, and T. J. Hope (2002)
Mol. Cell. Biol. 22, 2057-2067
   Abstract »    Full Text »    PDF »


ADVERTISEMENT
Click Me!

ADVERTISEMENT
Click Me!

To Advertise     Find Products

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