Limits...
Drosophila Pkaap regulates Rab4/Rab11-dependent traffic and Rab11 exocytosis of innate immune cargo.

Sorvina A, Shandala T, Brooks DA - Biol Open (2016)

Bottom Line: Here, we report that Drosophila Pkaap, an orthologue of the human dual-specific A-kinase-anchoring protein 2 or D-AKAP2 (also called AKAP10), appeared to have a nucleotide-dependent localisation to Rab4 and Rab11 endosomes.RNAi silencing of pkaap altered Rab4/Rab11 recycling endosome morphology, suggesting that Pkaap functions in cargo sorting and delivery in the secretory pathway.We propose that Pkaap has a dual role in antimicrobial peptide traffic and exocytosis, making it an essential component for the secretion of inflammatory mediators and the defence of the host against pathogens.

View Article: PubMed Central - PubMed

Affiliation: Sansom Institute for Health Research, University of South Australia, Adelaide, South Australia 5001, Australia.

No MeSH data available.


Related in: MedlinePlus

Pkaap colocates with Rab4 endosomes. (A-C) Confocal micrographs showing localisation of Pkaap detected with an anti-Pkaap antibody (red in A-C; greyscale in A′-C′) in relation to Rab4-YFP endosomes (green in A-C; greyscale in A″-C″) in fat body cells. Representative larval fat body cells were from the following genotypes: (A-A″; Rab4WT) wild-type control of Rab4, (B-B″; Rab4Q67L) GTP-bound constitutively active form of Rab4 and (C-C″; Rab4S22N) GDP-bound dominant negative form of Rab4. Scale bar: 5 μm. (D) Percentage of Pkaap colocation with Rab4-YFP vesicles in the fat body cells from Rab4WT, Rab4Q67L and Rab4S22N. One-way ANOVA and Tukey's multiple comparison test showed significant differences between the means for the genotypes (depicted by different letters on the bars, P<0.0001). Data presented as mean±s.e.m.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4920182&req=5

BIO016642F3: Pkaap colocates with Rab4 endosomes. (A-C) Confocal micrographs showing localisation of Pkaap detected with an anti-Pkaap antibody (red in A-C; greyscale in A′-C′) in relation to Rab4-YFP endosomes (green in A-C; greyscale in A″-C″) in fat body cells. Representative larval fat body cells were from the following genotypes: (A-A″; Rab4WT) wild-type control of Rab4, (B-B″; Rab4Q67L) GTP-bound constitutively active form of Rab4 and (C-C″; Rab4S22N) GDP-bound dominant negative form of Rab4. Scale bar: 5 μm. (D) Percentage of Pkaap colocation with Rab4-YFP vesicles in the fat body cells from Rab4WT, Rab4Q67L and Rab4S22N. One-way ANOVA and Tukey's multiple comparison test showed significant differences between the means for the genotypes (depicted by different letters on the bars, P<0.0001). Data presented as mean±s.e.m.

Mentions: In fat body cells from larvae expressing wild-type Rab4 (Rab4WT), Pkaap was detected on Rab4-YFP vesicles (Fig. 3A-A″) and in the cytosol (as previously observed for D-AKAP2; Eggers et al., 2009). In fat body cells expressing constitutively active Rab4Q67L, while the size of the Rab4 vesicles was reduced, Pkaap was still detected colocating with Rab4 vesicles (Fig. 3B-B″). In contrast, in fat body cells expressing dominant negative Rab4S22N, the size of the Rab4 vesicles was increased and Pkaap showed no colocation with these vesicles, but rather was only detected in the same proximity as these Rab4 endosomes (Fig. 3C-C″). Quantitative analysis showed that the expression of Rab4S22N significantly reduced Pkaap association with Rab4 vesicles, when compared to Rab4WT and Rab4Q67L (P<0.0001; Fig. 3D).Fig. 3.


Drosophila Pkaap regulates Rab4/Rab11-dependent traffic and Rab11 exocytosis of innate immune cargo.

Sorvina A, Shandala T, Brooks DA - Biol Open (2016)

Pkaap colocates with Rab4 endosomes. (A-C) Confocal micrographs showing localisation of Pkaap detected with an anti-Pkaap antibody (red in A-C; greyscale in A′-C′) in relation to Rab4-YFP endosomes (green in A-C; greyscale in A″-C″) in fat body cells. Representative larval fat body cells were from the following genotypes: (A-A″; Rab4WT) wild-type control of Rab4, (B-B″; Rab4Q67L) GTP-bound constitutively active form of Rab4 and (C-C″; Rab4S22N) GDP-bound dominant negative form of Rab4. Scale bar: 5 μm. (D) Percentage of Pkaap colocation with Rab4-YFP vesicles in the fat body cells from Rab4WT, Rab4Q67L and Rab4S22N. One-way ANOVA and Tukey's multiple comparison test showed significant differences between the means for the genotypes (depicted by different letters on the bars, P<0.0001). Data presented as mean±s.e.m.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4920182&req=5

BIO016642F3: Pkaap colocates with Rab4 endosomes. (A-C) Confocal micrographs showing localisation of Pkaap detected with an anti-Pkaap antibody (red in A-C; greyscale in A′-C′) in relation to Rab4-YFP endosomes (green in A-C; greyscale in A″-C″) in fat body cells. Representative larval fat body cells were from the following genotypes: (A-A″; Rab4WT) wild-type control of Rab4, (B-B″; Rab4Q67L) GTP-bound constitutively active form of Rab4 and (C-C″; Rab4S22N) GDP-bound dominant negative form of Rab4. Scale bar: 5 μm. (D) Percentage of Pkaap colocation with Rab4-YFP vesicles in the fat body cells from Rab4WT, Rab4Q67L and Rab4S22N. One-way ANOVA and Tukey's multiple comparison test showed significant differences between the means for the genotypes (depicted by different letters on the bars, P<0.0001). Data presented as mean±s.e.m.
Mentions: In fat body cells from larvae expressing wild-type Rab4 (Rab4WT), Pkaap was detected on Rab4-YFP vesicles (Fig. 3A-A″) and in the cytosol (as previously observed for D-AKAP2; Eggers et al., 2009). In fat body cells expressing constitutively active Rab4Q67L, while the size of the Rab4 vesicles was reduced, Pkaap was still detected colocating with Rab4 vesicles (Fig. 3B-B″). In contrast, in fat body cells expressing dominant negative Rab4S22N, the size of the Rab4 vesicles was increased and Pkaap showed no colocation with these vesicles, but rather was only detected in the same proximity as these Rab4 endosomes (Fig. 3C-C″). Quantitative analysis showed that the expression of Rab4S22N significantly reduced Pkaap association with Rab4 vesicles, when compared to Rab4WT and Rab4Q67L (P<0.0001; Fig. 3D).Fig. 3.

Bottom Line: Here, we report that Drosophila Pkaap, an orthologue of the human dual-specific A-kinase-anchoring protein 2 or D-AKAP2 (also called AKAP10), appeared to have a nucleotide-dependent localisation to Rab4 and Rab11 endosomes.RNAi silencing of pkaap altered Rab4/Rab11 recycling endosome morphology, suggesting that Pkaap functions in cargo sorting and delivery in the secretory pathway.We propose that Pkaap has a dual role in antimicrobial peptide traffic and exocytosis, making it an essential component for the secretion of inflammatory mediators and the defence of the host against pathogens.

View Article: PubMed Central - PubMed

Affiliation: Sansom Institute for Health Research, University of South Australia, Adelaide, South Australia 5001, Australia.

No MeSH data available.


Related in: MedlinePlus