Abstract

mRNA localization coupled with translational control is a widespread and conserved strategy that allows the localized production of proteins within eukaryotic cells. In Drosophila, oskar (osk) mRNA localization and translation at the posterior pole of the oocyte are essential for proper patterning of the embryo. Several P body components are involved in osk mRNA localization and translational repression, suggesting a link between P bodies and osk RNPs. In cultured mammalian cells, Ge-1 protein is required for P body formation. Combining genetic, biochemical and immunohistochemical approaches, we show that, in vivo, Drosophila Ge-1 (dGe-1) is an essential gene encoding a P body component that promotes formation of these structures in the germline. dGe-1 partially colocalizes with osk mRNA and is required for osk RNP integrity. Our analysis reveals that although under normal conditions dGe-1 function is not essential for osk mRNA localization, it becomes critical when other components of the localization machinery, such as staufen, Drosophila decapping protein 1 and barentsz are limiting. Our findings suggest an important role of dGe-1 in optimization of the osk mRNA localization process required for patterning the Drosophila embryo.

Highlights

  • MRNA localization coupled with translational control is a widespread and conserved strategy that allows the localized production of proteins within eukaryotic cells

  • Analysis of dGe-1D5 ovaries requires generation of homozygous mutant dGe-1D5 germline clones (GLC) [20]. Drosophila Ge-1 (dGe-1) mRNA levels are dramatically decreased in dGe-1D5 GLC ovaries and no trunfcated dGe-1 transcript produced from sequences downstream of the deleted region was detected (Figure S1B)

  • Western blots of extracts of young embryos probed with anti-dGe-1 antibody showed two bands of around 150 kDa, the upper of which was dramatically reduced in extracts of embryos derived from dGe-1D5 GLC, indicating that it represents dGe-1 protein (Figure 2A)

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Summary

Introduction

MRNA localization coupled with translational control is a widespread and conserved strategy that allows the localized production of proteins within eukaryotic cells. All RNAs tested, including the localized RNAs gurken (grk) (FlyBase: CG17610) [24], bicoid (bcd) (FlyBase: CG1034) [25] and osk [26,27], and the abundant RNAs, tubulin (tub) (FlyBase: CG8308) and ribosomal protein 49 (rp49) (FlyBase: CG7939), were enriched upon dGe-1 immunoprecipitation (Figure S2A), consistent with a general role of P bodies in mRNA regulation.

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