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An in vivo Crosslinking Approach to Isolate Protein Complexes From Drosophila Embryos
Published on: April 23, 2014
Drosophila germ granules are structured and contain homotypic mRNA clusters
Tatjana Trcek1, Markus Grosch1, Andrew York2
1Department of Cell Biology, HHMI, Skirball Institute of Biomolecular Medicine, NYU School of Medicine, 540 First Avenue, New York, New York 10016, USA.
Abstract:
Germ granules, specialized ribonucleoprotein particles, are a hallmark of all germ cells. In Drosophila, an estimated 200 mRNAs are enriched in the germ plasm, and some of these have important, often conserved roles in germ cell formation, specification, survival and migration. How mRNAs are spatially distributed within a germ granule and whether their position defines functional properties is unclear. Here we show, using single-molecule FISH and structured illumination microscopy, a super-resolution approach, that mRNAs are spatially organized within the granule whereas core germ plasm proteins are distributed evenly throughout the granule. Multiple copies of single mRNAs organize into 'homotypic clusters' that occupy defined positions within the center or periphery of the granule. This organization, which is maintained during embryogenesis and independent of the translational or degradation activity of mRNAs, reveals new regulatory mechanisms for germ plasm mRNAs that may be applicable to other mRNA granules.
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