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.

  • Letter
  • Published:

bicoid RNA localization requires specific binding of an endosomal sorting complex

Nature volume 445, pages 554–558 (2007)Cite this article

Abstract

bicoid messenger RNA localizes to the anterior of the Drosophila egg, where it is translated to form a morphogen gradient of Bicoid protein that patterns the head and thorax of the embryo. Although bicoid was the first localized cytoplasmic determinant to be identified1,2,3,4, little is known about how the mRNA is coupled to the microtubule-dependent transport pathway that targets it to the anterior, and it has been proposed that the mRNA is recognized by a complex of many redundant proteins, each of which binds to the localization element in the 3′ untranslated region (UTR) with little or no specificity5. Indeed, the only known RNA-binding protein that co-localizes with bicoid mRNA is Staufen, which binds non-specifically to double-stranded RNA in vitro6,7. Here we show that mutants in all subunits of the ESCRT-II complex (VPS22, VPS25 and VPS36) abolish the final Staufen-dependent step in bicoid mRNA localization. ESCRT-II is a highly conserved component of the pathway that sorts ubiquitinated endosomal proteins into internal vesicles8,9, and functions as a tumour-suppressor by removing activated receptors from the cytoplasm10,11. However, the role of ESCRT-II in bicoid localization seems to be independent of endosomal sorting, because mutations in ESCRT-I and III components do not affect the targeting of bicoid mRNA. Instead, VPS36 functions by binding directly and specifically to stem-loop V of the bicoid 3′ UTR through its amino-terminal GLUE domain12, making it the first example of a sequence-specific RNA-binding protein that recognizes the bicoid localization signal. Furthermore, VPS36 localizes to the anterior of the oocyte in a bicoid-mRNA-dependent manner, and is required for the subsequent recruitment of Staufen to the bicoid complex. This function of ESCRT-II as an RNA-binding complex is conserved in vertebrates and may clarify some of its roles that are independent of endosomal sorting.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Mutations in lsn and vps36 disrupt the anterior localization of GFP–Staufen and bicoid mRNA.
Figure 2: Localization of GFP–Stau in oocytes mutant for different ESCRT components.
Figure 3: VPS36 binds specifically to stem-loop V of the bicoid 3′ UTR.
Figure 4: VPS36 is recruited to the anterior of the oocyte by bicoid mRNA.

Similar content being viewed by others

References

  1. Frohnhöfer, H. G. & Nüsslein-Volhard, C. Organization of anterior pattern in the Drosophila embryo by the maternal gene bicoid. Nature 324, 120–125 (1986)

    Article  ADS  Google Scholar 

  2. Berleth, T. et al. The role of localization of bicoid RNA in organizing the anterior pattern of the Drosophila embryo. EMBO J. 7, 1749–1756 (1988)

    Article  CAS  Google Scholar 

  3. Driever, W. & Nüsslein-Volhard, C. A gradient of Bicoid protein in Drosophila embryos. Cell 54, 83–93 (1988)

    Article  CAS  Google Scholar 

  4. Driever, W. & Nüsslein-Volhard, C. The Bicoid protein determines position in the Drosophila embryo in a concentration-dependent manner. Cell 54, 95–104 (1988)

    Article  CAS  Google Scholar 

  5. Arn, E. A., Cha, B. J., Theurkauf, W. E. & Macdonald, P. M. Recognition of a bicoid mRNA localization signal by a protein complex containing Swallow, Nod, and RNA binding proteins. Dev. Cell 4, 41–51 (2003)

    Article  CAS  Google Scholar 

  6. St Johnston, D., Beuchle, D. & Nüsslein-Volhard, C. staufen, a gene required to localize maternal RNAs in the Drosophila egg. Cell 66, 51–63 (1991)

    Article  CAS  Google Scholar 

  7. Ramos, A. et al. RNA recognition by a Staufen double-stranded RNA-binding domain. EMBO J. 19, 997–1009 (2000)

    Article  CAS  Google Scholar 

  8. Babst, M. A protein's final ESCRT. Traffic 6, 2–9 (2005)

    Article  CAS  Google Scholar 

  9. Babst, M., Katzmann, D. J., Snyder, W. B., Wendland, B. & Emr, S. D. Endosome-associated complex, ESCRT-II, recruits transport machinery for protein sorting at the multivesicular body. Dev. Cell 3, 283–289 (2002)

    Article  CAS  Google Scholar 

  10. Vaccari, T. & Bilder, D. The Drosophila tumor suppressor vps25 prevents nonautonomous overproliferation by regulating Notch trafficking. Dev. Cell 9, 687–698 (2005)

    Article  CAS  Google Scholar 

  11. Thompson, B. J. et al. Tumor suppressor properties of the ESCRT-II complex component Vps25 in Drosophila. Dev. Cell 9, 711–720 (2005)

    Article  CAS  Google Scholar 

  12. Teo, H. et al. ESCRT-I core and ESCRT-II GLUE domain structures reveal role for GLUE in linking to ESCRT-I and membranes. Cell 125, 99–111 (2006)

    Article  CAS  Google Scholar 

  13. Frohnhöfer, H. G. & Nüsslein-Volhard, C. Maternal genes required for the anterior localization of bicoid activity in the embryo of Drosophila. Genes Dev. 1, 880–890 (1987)

    Article  Google Scholar 

  14. Macdonald, P. M., Luk, S. K. & Kilpatrick, M. Protein encoded by the exuperantia gene is concentrated at sites of bicoid mRNA accumulation in Drosophila nurse cells but not in oocytes or embryos. Genes Dev. 5, 2455–2466 (1991)

    Article  CAS  Google Scholar 

  15. Wilhelm, J. E. et al. Isolation of a ribonucleoprotein complex involved in mRNA localization in Drosophila oocytes. J. Cell Biol. 148, 427–440 (2000)

    Article  CAS  Google Scholar 

  16. Theurkauf, W. E. & Hazelrigg, T. I. In vivo analyses of cytoplasmic transport and cytoskeletal organization during Drosophila oogenesis: characterization of a multi-step anterior localization pathway. Development 125, 3655–3666 (1998)

    CAS  PubMed  Google Scholar 

  17. Moon, W. & Hazelrigg, T. The Drosophila microtubule-associated protein Mini spindles is required for cytoplasmic microtubules in oogenesis. Curr. Biol. 14, 1957–1961 (2004)

    Article  CAS  Google Scholar 

  18. Vogt, N., Koch, I., Schwarz, H., Schnorrer, F. & Nüsslein-Volhard, C. The γTuRC components Grip75 and Grip128 have an essential microtubule-anchoring function in the Drosophila germline. Development 133, 3963–3972 (2006)

    Article  CAS  Google Scholar 

  19. Ferrandon, D., Elphick, L., Nüsslein-Volhard, C. & St Johnston, D. Staufen protein associates with the 3′UTR of bicoid mRNA to form particles that move in a microtubule-dependent manner. Cell 79, 1221–1232 (1994)

    Article  CAS  Google Scholar 

  20. Weil, T. T., Forrest, K. M. & Gavis, E. R. Localization of bicoid mRNA in late oocytes is maintained by continual active transport. Dev. Cell 11, 251–262 (2006)

    Article  CAS  Google Scholar 

  21. St Johnston, D., Driever, W., Berleth, T., Richstein, S. & Nüsslein-Volhard, C. Multiple steps in the localization of bicoid RNA to the anterior pole of the Drosophila oocyte. Development 107, (Suppl)13–19 (1989)

    CAS  PubMed  Google Scholar 

  22. Macdonald, P. M., Kerr, K., Smith, J. L. & Leask, A. RNA regulatory element BLE1 directs the early steps of bicoid mRNA localization. Development 118, 1233–1243 (1993)

    CAS  PubMed  Google Scholar 

  23. Micklem, D. R., Adams, J., Grunert, S. & St Johnston, D. Distinct roles of two conserved Staufen domains in oskar mRNA localization and translation. EMBO J. 19, 1366–1377 (2000)

    Article  CAS  Google Scholar 

  24. Martin, S. G., Leclerc, V., Smith-Litière, K. & St Johnston, D. The identification of novel genes required for Drosophila anteroposterior axis formation in a germline clone screen using GFP-Staufen. Development 130, 4201–4215 (2003)

    Article  CAS  Google Scholar 

  25. Teo, H., Perisic, O., Gonzalez, B. & Williams, R. L. ESCRT-II, an endosome-associated complex required for protein sorting: crystal structure and interactions with ESCRT-III and membranes. Dev. Cell 7, 559–569 (2004)

    Article  CAS  Google Scholar 

  26. Moberg, K. H., Schelble, S., Burdick, S. K. & Hariharan, I. K. Mutations in erupted, the Drosophila ortholog of mammalian Tumor Susceptibility Gene 101, elicit non-cell-autonomous overgrowth. Dev. Cell 9, 699–710 (2005)

    Article  CAS  Google Scholar 

  27. Kamura, T. et al. Cloning and characterization of ELL-associated proteins EAP45 and EAP20. A role for yeast EAP-like proteins in regulation of gene expression by glucose. J. Biol. Chem. 276, 16528–16533 (2001)

    Article  CAS  Google Scholar 

  28. Schmidt, A. E., Miller, T., Schmidt, S. L., Shiekhattar, R. & Shilatifard, A. Cloning and characterization of the EAP30 subunit of the ELL complex that confers derepression of transcription by RNA polymerase II. J. Biol. Chem. 274, 21981–21985 (1999)

    Article  CAS  Google Scholar 

  29. Jin, Y., Mancuso, J. J., Uzawa, S., Cronembold, D. & Cande, W. Z. The fission yeast homolog of the human transcription factor EAP30 blocks meiotic spindle pole body amplification. Dev. Cell 9, 63–73 (2005)

    Article  CAS  Google Scholar 

  30. Putz, U., Skehel, P. & Kuhl, D. A tri-hybrid system for the analysis and detection of RNA–protein interactions. Nucleic Acids Res. 24, 4838–4840 (1996)

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We are very grateful to R. Williams, O. Perisec and H. Teo for giving us the purified recombinant Xenopus laevis VPS36 GLUE domain, and for their helpful advice on ESCRT-II. We thank J. Oberdick and Z. Zhang for sharing their unpublished results on rat VPS36. This work was supported by a Wellcome Trust Principal Research Fellowship to D.St J. and by the Max-Planck-Gesellschaft (U.I.).

Author information

Author notes

  1. Uwe Irion

    Present address: Max-Planck-Institut für Entwicklungsbiologie, Spemannstraße 35, 72076, Tübingen, Germany

Authors and Affiliations

  1. Wellcome Trust/Cancer Research UK Gurdon Institute and Department of Genetics, University of Cambridge, Tennis Court Road, Cambridge, CB2 1QN, UK

    Uwe Irion & Daniel St Johnston

Authors
  1. Uwe Irion
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Daniel St Johnston
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Daniel St Johnston.

Ethics declarations

Competing interests

GenBank accession numbers for the VPS36 genomic rescue construct and the Venus–VPS36 construct (see Supplementary Information) are EF152770 and EF152771, respectively. Reprints and permissions information is available at www.nature.com/reprints. The authors declare no competing financial interests.

Supplementary information

Supplementary Information

This file contains Supplementary Methods, Supplementary Figures S1-S2 and additional references. (PDF 1272 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Irion, U., St Johnston, D. bicoid RNA localization requires specific binding of an endosomal sorting complex. Nature 445, 554–558 (2007). https://doi.org/10.1038/nature05503

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nature05503

This article is cited by

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

Advanced 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