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Drosophila Ge-1 promotes P body formation and oskar mRNA localization.

Fan SJ, Marchand V, Ephrussi A - PLoS ONE (2011)

Bottom Line: 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.Our findings suggest an important role of dGe-1 in optimization of the osk mRNA localization process required for patterning the Drosophila embryo.

View Article: PubMed Central - PubMed

Affiliation: Developmental Biology Unit, European Molecular Biology Laboratory, Heidelberg, Germany.

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.

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dGe-1 cooperates with Stau and dDcp1 in osk mRNA localization and embryonic patterning.(A–I) Simultaneous detection of osk mRNA (red) and protein (green) by FISH coupled with immunodetection using a rabbit anti-Osk antibody. osk mRNA (A) and protein (B) colocalize in a posterior crescent at the posterior pole of wt oocytes at S10. (D–I) Examples of the aberrant distribution of osk mRNA and protein in dGe-1Δ5 GLC egg-chambers. Overlays of osk mRNA and Osk protein signals (C, F, I). DNA stained with 4,6-diamidino-2-phenylindole (DAPI) (blue). (J) Quantification (%) of osk mRNA localization patterns in S10 egg-chambers in different genetic backgrounds. Normal and abnormal osk mRNA localization are represented as black and grey bars, respectively. n represents the number of embryos analyzed. (K–M) Loss of maternal dGe-1 causes aberrant cuticle patterning in some embryos derived from dGe-1Δ5 GLC. Lateral view of embryos oriented anterior to the left, ventral side down. (K) In a wt embryo, posterior structures represented by the abdominal denticle belts are clearly visible along the ventral side. In a portion of embryos derived from dGe-1Δ5 GLC, several denticle belts are missing (L) and, in extreme cases, all are absent (M). (N) Quantification of posterior group phenotypes produced in different genetic backgrounds. Embryos lacking at least one (weak) or all (strong) abdominal denticle belts were quantified. n represents the number of embryos analyzed. Bar, 50 µm.
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pone-0020612-g003: dGe-1 cooperates with Stau and dDcp1 in osk mRNA localization and embryonic patterning.(A–I) Simultaneous detection of osk mRNA (red) and protein (green) by FISH coupled with immunodetection using a rabbit anti-Osk antibody. osk mRNA (A) and protein (B) colocalize in a posterior crescent at the posterior pole of wt oocytes at S10. (D–I) Examples of the aberrant distribution of osk mRNA and protein in dGe-1Δ5 GLC egg-chambers. Overlays of osk mRNA and Osk protein signals (C, F, I). DNA stained with 4,6-diamidino-2-phenylindole (DAPI) (blue). (J) Quantification (%) of osk mRNA localization patterns in S10 egg-chambers in different genetic backgrounds. Normal and abnormal osk mRNA localization are represented as black and grey bars, respectively. n represents the number of embryos analyzed. (K–M) Loss of maternal dGe-1 causes aberrant cuticle patterning in some embryos derived from dGe-1Δ5 GLC. Lateral view of embryos oriented anterior to the left, ventral side down. (K) In a wt embryo, posterior structures represented by the abdominal denticle belts are clearly visible along the ventral side. In a portion of embryos derived from dGe-1Δ5 GLC, several denticle belts are missing (L) and, in extreme cases, all are absent (M). (N) Quantification of posterior group phenotypes produced in different genetic backgrounds. Embryos lacking at least one (weak) or all (strong) abdominal denticle belts were quantified. n represents the number of embryos analyzed. Bar, 50 µm.

Mentions: In addition, in contrast to wt oocytes (Figure 3A–C), a significant proportion of dGe-1Δ5 GLC oocytes displays defective osk mRNA localization at the posterior pole at stages 9 and 10 (S9, S10) (Figure 3D, 3G and 3J and Figure S3A). This mislocalization, which is rescued by transgenic expression of a dGe-1-B cDNA (Figure S3B–D), is paralleled by defects in Osk protein localization (Figure 3E and 3H), and Osk protein levels are reduced in dGe-1Δ5 GLC ovaries (Figure S3E).


Drosophila Ge-1 promotes P body formation and oskar mRNA localization.

Fan SJ, Marchand V, Ephrussi A - PLoS ONE (2011)

dGe-1 cooperates with Stau and dDcp1 in osk mRNA localization and embryonic patterning.(A–I) Simultaneous detection of osk mRNA (red) and protein (green) by FISH coupled with immunodetection using a rabbit anti-Osk antibody. osk mRNA (A) and protein (B) colocalize in a posterior crescent at the posterior pole of wt oocytes at S10. (D–I) Examples of the aberrant distribution of osk mRNA and protein in dGe-1Δ5 GLC egg-chambers. Overlays of osk mRNA and Osk protein signals (C, F, I). DNA stained with 4,6-diamidino-2-phenylindole (DAPI) (blue). (J) Quantification (%) of osk mRNA localization patterns in S10 egg-chambers in different genetic backgrounds. Normal and abnormal osk mRNA localization are represented as black and grey bars, respectively. n represents the number of embryos analyzed. (K–M) Loss of maternal dGe-1 causes aberrant cuticle patterning in some embryos derived from dGe-1Δ5 GLC. Lateral view of embryos oriented anterior to the left, ventral side down. (K) In a wt embryo, posterior structures represented by the abdominal denticle belts are clearly visible along the ventral side. In a portion of embryos derived from dGe-1Δ5 GLC, several denticle belts are missing (L) and, in extreme cases, all are absent (M). (N) Quantification of posterior group phenotypes produced in different genetic backgrounds. Embryos lacking at least one (weak) or all (strong) abdominal denticle belts were quantified. n represents the number of embryos analyzed. Bar, 50 µm.
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pone-0020612-g003: dGe-1 cooperates with Stau and dDcp1 in osk mRNA localization and embryonic patterning.(A–I) Simultaneous detection of osk mRNA (red) and protein (green) by FISH coupled with immunodetection using a rabbit anti-Osk antibody. osk mRNA (A) and protein (B) colocalize in a posterior crescent at the posterior pole of wt oocytes at S10. (D–I) Examples of the aberrant distribution of osk mRNA and protein in dGe-1Δ5 GLC egg-chambers. Overlays of osk mRNA and Osk protein signals (C, F, I). DNA stained with 4,6-diamidino-2-phenylindole (DAPI) (blue). (J) Quantification (%) of osk mRNA localization patterns in S10 egg-chambers in different genetic backgrounds. Normal and abnormal osk mRNA localization are represented as black and grey bars, respectively. n represents the number of embryos analyzed. (K–M) Loss of maternal dGe-1 causes aberrant cuticle patterning in some embryos derived from dGe-1Δ5 GLC. Lateral view of embryos oriented anterior to the left, ventral side down. (K) In a wt embryo, posterior structures represented by the abdominal denticle belts are clearly visible along the ventral side. In a portion of embryos derived from dGe-1Δ5 GLC, several denticle belts are missing (L) and, in extreme cases, all are absent (M). (N) Quantification of posterior group phenotypes produced in different genetic backgrounds. Embryos lacking at least one (weak) or all (strong) abdominal denticle belts were quantified. n represents the number of embryos analyzed. Bar, 50 µm.
Mentions: In addition, in contrast to wt oocytes (Figure 3A–C), a significant proportion of dGe-1Δ5 GLC oocytes displays defective osk mRNA localization at the posterior pole at stages 9 and 10 (S9, S10) (Figure 3D, 3G and 3J and Figure S3A). This mislocalization, which is rescued by transgenic expression of a dGe-1-B cDNA (Figure S3B–D), is paralleled by defects in Osk protein localization (Figure 3E and 3H), and Osk protein levels are reduced in dGe-1Δ5 GLC ovaries (Figure S3E).

Bottom Line: 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.Our findings suggest an important role of dGe-1 in optimization of the osk mRNA localization process required for patterning the Drosophila embryo.

View Article: PubMed Central - PubMed

Affiliation: Developmental Biology Unit, European Molecular Biology Laboratory, Heidelberg, Germany.

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.

Show MeSH
Related in: MedlinePlus