Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

CRM1 is responsible for intracellular transport mediated by the nuclear export signal

Abstract

The discovery of nuclear export signals (NESs) in a number of proteins revealed the occurrence of signal-dependent transport of proteins from the nucleus to the cytoplasm1,2,3,4,5,6,7,8,9,10,11,12,13,14. Although the consensus motif of the NESs has been shown to be a leucine-rich, short amino-acid sequence2,6,7, its receptor has not been identified. A cytotoxin leptomycin B (LMB) has recently been suggested to inhibit the NES-mediated transport of Rev protein15. Here we show that LMB is a potent and specific inhibitor of the NES-dependent nuclear export of proteins. Moreover, we have found a protein of relative molecular mass 110K (p110) in Xenopus oocyte extracts that binds to the intact NES but not to the mutated, non-functional NES. The binding of p110 to NES is inhibited by LMB. We show that p110 is CRM1, which is an evolutionarily conserved protein16,17,18 originally found as an essential nuclear protein in fission yeast16 and known as a likely target of LMB19. We also show that nuclear export of a fission yeast protein, Dsk1, which has a leucine-rich NES, is disrupted in wild-type yeast treated with LMB or in the crm1 mutant. These results indicate that CRM1 is an essential mediator of the NES-dependent nuclear export of proteins in eukaryotic cells.

This is a preview of subscription content

Access options

Buy article

Get time limited or full article access on ReadCube.

$32.00

All prices are NET prices.

Figure 1: LMB is an inhibitor of the NES-dependent nuclear export of proteins.
Figure 2: The p110 protein binds specifically to NES in an LMB-sensitive manner.
Figure 3: Disruption of cytoplasmic localization and nuclear exclusion of Dsk1 in a fission-yeast crm1 mutant.

References

  1. Fischer, U., Huber, J., Boelens, W. C., Mattaj, I. W. & Lührmann, R. The HIV-1 Rev activation domain is a nuclear export signal that accesses an export pathway used by specific cellular RNAs. Cell 82, 475–483 (1995).

    CAS  Article  Google Scholar 

  2. Wen, W., Meinkoth, J. L., Tsien, R. Y. & Taylor, S. S. Identification of a signal for rapid export of proteins from the nucleus. Cell 82, 463–473 (1995).

    CAS  Article  Google Scholar 

  3. Fridell, R. A. et al. Amphibian transcription factor IIIA proteins contain a sequence element functionally equivalent to the nuclear export signal of human immunodeficiency virus type 1 Rev. Proc. Natl Acad. Sci. USA 93, 2936–2940 (1996).

    ADS  CAS  Article  Google Scholar 

  4. Meyer, B. E., Meinkoth, J. L. & Malim, M. H. Nuclear transport of human immunodeficiency virus type 1, visna virus, and equine infectious anemia virus Rev proteins: identification of a family of transferable nuclear export signals. J. Virol. 70, 2350–2359 (1996).

    CAS  PubMed  PubMed Central  Google Scholar 

  5. Fukuda, M., Gotoh, I., Gotoh, Y. & Nishida, E. Cytoplasmic localizaiton of MAP kinase kinase directed by its N-terminal, leucine-rich short amino acid sequence, which acts as a nuclear export signal. J. Biol. Chem. 271, 20024–20028 (1996).

    CAS  Article  Google Scholar 

  6. Bogerd, H. P., Fridell, R. A., Benson, R. E., Hua, J. & Cullen, B. R. Protein sequence requirements for function of the human T-cell leukemia virus type 1 Rex nuclear export signal delineated by a novel in vivo randomization-selection assay. Mol. Cell. Biol. 16, 4207–4214 (1996).

    CAS  Article  Google Scholar 

  7. Kim, F. J., Beeche, A. A., Hunter, J. J., Chin, D. J. & Hope, T. J. Characterization of the nuclear export signal of human T-cell lymphotropic virus type 1 Rex reveals that nuclear export is mediated by position-varaible hydrophobic interactions. Mol. Cell. Biol. 16, 5147–5155 (1996).

    CAS  Article  Google Scholar 

  8. Fridell, R. A., Benson, R. E., Hua, J., Bogerd, H. P. & Cullen, B. R. Anuclear role for the Fragile X mental retardation protein. EMBO J. 15, 5408–5414 (1996).

    CAS  Article  Google Scholar 

  9. Richards, S. A., Lounsbury, K. M., Carey, K. L. & Macara, I. G. Anuclear export signal is essential for the cytosolic localization of the Ran binding protein, RanBP1. J. Cell Biol. 134, 1157–1168 (1996).

    CAS  Article  Google Scholar 

  10. Murphy, R. & Wente, S. R. An RNA-export mediator with an essential nuclear export signal. Nature 383, 357–360 (1996).

    ADS  CAS  Article  Google Scholar 

  11. Gerace, L. Nuclear export signals and the fast track to the cytoplasm. Cell 82, 341–344 (1995).

    CAS  Article  Google Scholar 

  12. Görlich, D. & Mattaj, I. W. Nucleocytoplasmic transport. Science 271, 1513–1518 (1996).

    ADS  Article  Google Scholar 

  13. Nigg, E. A. Nucleocytoplasmic transport: signals, mechanisms and regulation. Nature 386, 779–787 (1997).

    ADS  CAS  Article  Google Scholar 

  14. Nakielny, S. & Dreyfuss, G. Nuclear export of proteins and RNAs. Curr. Opin. Cell Biol. 9, 420–429 (1997).

    CAS  Article  Google Scholar 

  15. Wolff, B., Sanglier, J. J. & Wang, Y. Leptomycin B is an inhibitor of nuclear export: inhibition of nucleo-cytoplasmic translocation of the human immunodeficiency virus type 1 (HIV-1) Rev protein and Rev-dependent mRNA. Chem. Biol. 4, 139–147 (1997).

    CAS  Article  Google Scholar 

  16. Adachi, Y. & Yanagida, M. Higher order chromosome structure is affected by cold-sensitive mutations in a Schizosaccharomyces pombegene crm1+ which encodes a 115-kD protein preferentially localized in the nucleus and at its periphery. J. Cell Biol. 108, 1195–1207 (1989).

    CAS  Article  Google Scholar 

  17. Toda, T. et al. Fission yeast pap1-dependent transcription is negatively regulated by an essential nuclear protein, crm1. Mol. Cell. Biol. 12, 5474–5484 (1992).

    CAS  Article  Google Scholar 

  18. Fornerod, M. et al. The human homologue of yeast CRM1 is in a dynamic subcomplex with CAN/Nup214 and a novel nuclear pore component Nup88. EMBO J. 16, 807–816 (1997).

    CAS  Article  Google Scholar 

  19. Nishi, K. et al. Leptomycin B targets a regulatory cascade of crm1, a fission yeast nuclear protein, involved in control of higher order chromosome structure and gene expression. J. Biol. Chem. 269, 6320–6324 (1994).

    CAS  PubMed  Google Scholar 

  20. Fornerod, M., Boer, J., van Baal, S., Morreau, H. & Grosveld, G. Interaction of cellular proteins with the leukemia specific fusion proteins DEK-AN and SET-CAN and their normal coutnerpart, the nucleoporin CAN. Oncogene 13, 1801–1808 (1996).

    CAS  PubMed  Google Scholar 

  21. Gui, J. F., Lane, W. S. & Fu, X-D. Aserine kinase regulates intracellular localization of splicing factors in the cell cycle. Nature 369, 678–682 (1994).

    ADS  CAS  Article  Google Scholar 

  22. Takeuchi, M. & Yanagida, M. Amitotic role for a novel fission yeast protein kinase dsk1 with cell cycle stage dependent phosphorylation and localization. Mol. Biol. Cell 4, 247–260 (1993).

    CAS  Article  Google Scholar 

  23. Stuzt, 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 82, 495–506 (1995).

    Article  Google Scholar 

  24. Bogerd, H. P., Fridell, R. A., Madore, S. & Cullen, B. R. Identification of a novel cellular cofactor for the Rev/Rex class of retoroviral regulatory proteins. Cell 82, 485–494 (1995).

    CAS  Article  Google Scholar 

  25. Fritz, C. C., Zapp, M. L. & Green, M. R. Ahuman nucleoporin-like protein that specifically interacts with HIV Rev. Nature 376, 530–533 (1995).

    ADS  CAS  Article  Google Scholar 

  26. Fukuda, M., Gotoh, Y. & Nishida, E. Interaction of MAP kinase with MAP kinase kinase: Its possible role in the control of nucleocytoplasmic transport of MAP kinase. EMBO J. 16, 1901–1908 (1997).

    CAS  Article  Google Scholar 

  27. Stade, K., Ford, C. S., Guthrie, C. & Weis, K. Exportin 1 (Crm1p) is an essential nuclear export factor. Cell 90, 1041–1050 (1997).

    CAS  Article  Google Scholar 

  28. Fornerod, M., Ohno, M., Yoshida, M. & Maltay, I. W. CRM1 is an export receptor for leucine-rich nuclear export signals. Cell 90, 1051–1055 (1997).

    CAS  Article  Google Scholar 

  29. Ossareh-Nazari, B., Bachelerie, F. & Dargemont, C. Evidence for a role of CRM1 in signal-mediated nuclear protein export. Science 278, 141–144 (1997).

    CAS  Article  Google Scholar 

Download references

Acknowledgements

We thank K. Kumada for comments and discussion, and M. Watanabe for preparation of Dsk1-NES-OVA. This work was supported by grants from the Ministry of Education, Science and Culture of Japan (E.N.) and the Japan Science Technology Corporation (M. Yanagida).

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Eisuke Nishida.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Fukuda, M., Asano, S., Nakamura, T. et al. CRM1 is responsible for intracellular transport mediated by the nuclear export signal. Nature 390, 308–311 (1997). https://doi.org/10.1038/36894

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/36894

Further reading

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing