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.
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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 We thank J. Oberdick and Z. Zhang for sharing their unpublished results on rat VPS36.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.).
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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.
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This file contains Supplementary Methods, Supplementary Figures S1-S2 and additional references. (PDF 1272 kb)
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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
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DOI: https://doi.org/10.1038/nature05503
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