Limits...
A novel role for dp115 in the organization of tER sites in Drosophila.

Kondylis V, Rabouille C - J. Cell Biol. (2003)

Bottom Line: However, we found that this morphologically altered exocytic pathway was nevertheless largely competent in anterograde protein transport using two different assays.The effects were specific for dp115.Depletion of the Drosophila homologues of GM130 and syntaxin 5 (dSed5p) did not lead to an effect on the tER organization, though the Golgi stacks were greatly vesiculated in the cells depleted of dSed5p.

View Article: PubMed Central - PubMed

Affiliation: The Wellcome Trust Center for Cell Biology, Institute for Cell and Molecular Biology, University of Edinburgh, UK.

ABSTRACT
Here, we describe that depletion of the Drosophila homologue of p115 (dp115) by RNA interference in Drosophila S2 cells led to important morphological changes in the Golgi stack morphology and the transitional ER (tER) organization. Using conventional and immunoelectron microscopy and confocal immunofluorescence microscopy, we show that Golgi stacks were converted into clusters of vesicles and tubules, and that the tERs (marked by Sec23p) lost their focused organization and were now dispersed throughout the cytoplasm. However, we found that this morphologically altered exocytic pathway was nevertheless largely competent in anterograde protein transport using two different assays. The effects were specific for dp115. Depletion of the Drosophila homologues of GM130 and syntaxin 5 (dSed5p) did not lead to an effect on the tER organization, though the Golgi stacks were greatly vesiculated in the cells depleted of dSed5p. Taken together, these studies suggest that dp115 could be implicated in the architecture of both the Golgi stacks and the tER sites.

Show MeSH

Related in: MedlinePlus

Effect of depleting dp115 and dGM130 on the Golgi stack mor-phology. Drosophila S2 cells were cultured (A) in the absence (−dsRNA) or (B) presence of ds dGM130 for 96 h, or (C) in the presence of ds dp115 for 72 or (D) 96 h. Cells were collected and processed for conventional EM. Golgi stacks are indicated with a long arrow, and clusters of vesicles and tubules are marked between brackets. N, nucleus. Bars, 200 nm.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC2172793&req=5

fig3: Effect of depleting dp115 and dGM130 on the Golgi stack mor-phology. Drosophila S2 cells were cultured (A) in the absence (−dsRNA) or (B) presence of ds dGM130 for 96 h, or (C) in the presence of ds dp115 for 72 or (D) 96 h. Cells were collected and processed for conventional EM. Golgi stacks are indicated with a long arrow, and clusters of vesicles and tubules are marked between brackets. N, nucleus. Bars, 200 nm.

Mentions: We depleted S2 cells of dGM130 using a dsRNA corresponding to the second exon of dGM130 (ds dGM130). When samples were analyzed by Western blotting, two bands were detected, one very faint, which was neglected (Fig. 2, top, lane C), and a strong one. The strong band migrated at the predicted position for a protein of the mass of dGM130 (arrow) and was reduced below detectable level after 48 h (Fig. 2, top) and remained so up to 120 h of incubation. This depletion, however, did not lead to any effect on Golgi stack morphology, as assessed by EM (Fig. 3 B).


A novel role for dp115 in the organization of tER sites in Drosophila.

Kondylis V, Rabouille C - J. Cell Biol. (2003)

Effect of depleting dp115 and dGM130 on the Golgi stack mor-phology. Drosophila S2 cells were cultured (A) in the absence (−dsRNA) or (B) presence of ds dGM130 for 96 h, or (C) in the presence of ds dp115 for 72 or (D) 96 h. Cells were collected and processed for conventional EM. Golgi stacks are indicated with a long arrow, and clusters of vesicles and tubules are marked between brackets. N, nucleus. Bars, 200 nm.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2172793&req=5

fig3: Effect of depleting dp115 and dGM130 on the Golgi stack mor-phology. Drosophila S2 cells were cultured (A) in the absence (−dsRNA) or (B) presence of ds dGM130 for 96 h, or (C) in the presence of ds dp115 for 72 or (D) 96 h. Cells were collected and processed for conventional EM. Golgi stacks are indicated with a long arrow, and clusters of vesicles and tubules are marked between brackets. N, nucleus. Bars, 200 nm.
Mentions: We depleted S2 cells of dGM130 using a dsRNA corresponding to the second exon of dGM130 (ds dGM130). When samples were analyzed by Western blotting, two bands were detected, one very faint, which was neglected (Fig. 2, top, lane C), and a strong one. The strong band migrated at the predicted position for a protein of the mass of dGM130 (arrow) and was reduced below detectable level after 48 h (Fig. 2, top) and remained so up to 120 h of incubation. This depletion, however, did not lead to any effect on Golgi stack morphology, as assessed by EM (Fig. 3 B).

Bottom Line: However, we found that this morphologically altered exocytic pathway was nevertheless largely competent in anterograde protein transport using two different assays.The effects were specific for dp115.Depletion of the Drosophila homologues of GM130 and syntaxin 5 (dSed5p) did not lead to an effect on the tER organization, though the Golgi stacks were greatly vesiculated in the cells depleted of dSed5p.

View Article: PubMed Central - PubMed

Affiliation: The Wellcome Trust Center for Cell Biology, Institute for Cell and Molecular Biology, University of Edinburgh, UK.

ABSTRACT
Here, we describe that depletion of the Drosophila homologue of p115 (dp115) by RNA interference in Drosophila S2 cells led to important morphological changes in the Golgi stack morphology and the transitional ER (tER) organization. Using conventional and immunoelectron microscopy and confocal immunofluorescence microscopy, we show that Golgi stacks were converted into clusters of vesicles and tubules, and that the tERs (marked by Sec23p) lost their focused organization and were now dispersed throughout the cytoplasm. However, we found that this morphologically altered exocytic pathway was nevertheless largely competent in anterograde protein transport using two different assays. The effects were specific for dp115. Depletion of the Drosophila homologues of GM130 and syntaxin 5 (dSed5p) did not lead to an effect on the tER organization, though the Golgi stacks were greatly vesiculated in the cells depleted of dSed5p. Taken together, these studies suggest that dp115 could be implicated in the architecture of both the Golgi stacks and the tER sites.

Show MeSH
Related in: MedlinePlus