Article | Published:

Independent and coordinate trafficking of single Drosophila germ plasm mRNAs

Nature Cell Biology volume 17, pages 558568 (2015) | Download Citation

  • A Corrigendum to this article was published on 25 February 2016

This article has been updated

Abstract

Messenger RNA localization is a conserved mechanism for spatial control of protein synthesis, with key roles in generating cellular and developmental asymmetry. Whereas different transcripts may be targeted to the same subcellular domain, the extent to which their localization is coordinated is unclear. Using quantitative single-molecule imaging, we analysed the assembly of Drosophila germ plasm mRNA granules inherited by nascent germ cells. We find that the germ-cell-destined transcripts nanos, cyclin B and polar granule component travel within the oocyte as ribonucleoprotein particles containing single mRNA molecules but co-assemble into multi-copy heterogeneous granules selectively at the posterior of the oocyte. The stoichiometry and dynamics of assembly indicate a defined stepwise sequence. Our data suggest that co-packaging of these transcripts ensures their effective segregation to germ cells. In contrast, compartmentalization of the germline determinant oskar mRNA into different granules limits its entry into germ cells. This exclusion is required for proper germline development.

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Change history

  • 01 February 2016

    In the version of this Article originally published, the sentence ‘13% of GFP–Vas granules contain nos mRNAs and 11% contain cycB’ in the caption of Fig. 4h–j was incorrect; it should have read ‘69% of GFP–Vas granules contain nos and 51% contain cycB’. This has been corrected in all online versions of the Article.

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Acknowledgements

We thank W. Eagle (Princeton University, USA), P. Lasko (McGill University, Canada), P. Macdonald (University of Texas, Austin, USA) and D. St Johnston (Gurdon Institute, UK) for fly stocks and reagents, S. Chatterjee and S. Kyin for technical assistance, and E. Abbaszadeh, S. Blythe and B. He for comments on the manuscript. This work was financially supported by National Institute of Health grant R01GM067758 (E.R.G.) and the Howard Hughes Medical Institute (E.F.W.).

Author information

Affiliations

  1. Department of Molecular Biology, Princeton University, Princeton, New Jersey 08544, USA

    • Shawn C. Little
    • , Kristina S. Sinsimer
    • , Jack J. Lee
    • , Eric F. Wieschaus
    •  & Elizabeth R. Gavis
  2. Howard Hughes Medical Institute, Princeton University, Princeton, New Jersey 08544, USA

    • Shawn C. Little
    •  & Eric F. Wieschaus

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Contributions

S.C.L., K.S.S. and E.R.G. designed the experiments. S.C.L., K.S.S., J.J.L. and E.R.G. performed the experiments S.C.L. and E.R.G. analysed the data. S.C.L., E.R.G. and E.F.W. wrote the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Elizabeth R. Gavis.

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Videos

  1. 1.

    Subsection of a confocal stack of an early (n.c. 3) embryo posterior labeled with nos probes.

    First section is adjacent to the lateral cortex. Stack is shown twice, followed by magnified views of unlocalized and localized regions. To facilitate visualization of unlocalized punctae, the stack is displayed at high contrast such that localized granules appear saturated. However, no saturated pixels are present in raw images, as illustrated with a single cortical image slice displayed at alternating high and low contrast settings. Z-sections are separated by 340 nm.

  2. 2.

    Time lapse movie of germ plasm RNP particles containing nos*GFP at the oocyte posterior.

    Maximum projection of 5 z-sections spanning a total of 6 μm and a total time of 5 min.

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DOI

https://doi.org/10.1038/ncb3143

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