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De novo actin polymerization is required for model Hirano body formation in Dictyostelium.

Dong Y, Shahid-Salles S, Sherling D, Fechheimer N, Iyer N, Wells L, Fechheimer M, Furukawa R - Biol Open (2016)

Bottom Line: Whereas model Hirano bodies could form in WASH-deficient cells, they failed to form in cells lacking HSPC300, a member of the WAVE complex.In the case of VASP, both its G- and F-actin binding domains were required for model Hirano body formation.Collectively, our results indicate that de novo actin polymerization is required to form model Hirano bodies.

View Article: PubMed Central - PubMed

Affiliation: Department of Cellular Biology, University of Georgia, Athens, GA, USA 30602.

No MeSH data available.


Related in: MedlinePlus

Model Hirano bodies form after removal of CK666. Cells were induced to express E60K-GFP and immediately incubated with CK666 for 12 h. They recovered in media after CK666 was removed for another 12 h (left column). Cells that were induced to express E60K-GFP at the same time (middle column) and at the time of CK666 removal (right column) were also observed as controls. All cells were examined by DIC (first row), and by fluorescence microscopy using TRITC-phalloidin (second row) or GFP (third row). Model Hirano bodies formed in a substantial amount of cells in all three samples. Scale bar=15 µm.
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BIO014944F5: Model Hirano bodies form after removal of CK666. Cells were induced to express E60K-GFP and immediately incubated with CK666 for 12 h. They recovered in media after CK666 was removed for another 12 h (left column). Cells that were induced to express E60K-GFP at the same time (middle column) and at the time of CK666 removal (right column) were also observed as controls. All cells were examined by DIC (first row), and by fluorescence microscopy using TRITC-phalloidin (second row) or GFP (third row). Model Hirano bodies formed in a substantial amount of cells in all three samples. Scale bar=15 µm.

Mentions: To rule out the possibility that the inhibition of model Hirano body formation by CK666 was due to a non-specific inhibition, we removed CK666 after 12 h of incubation and allowed the cells to recover for another 12 h with media under conditions for expression of E60K-GFP. Following removal of CK666, model Hirano bodies formed in a significant proportion of cells (Fig. 5). Thus, the effect of CK666 is reversible, and the process of model Hirano body formation can still be triggered. Therefore CK666, which inhibits the Arp2/3 complex activity, also inhibits model Hirano body formation and confirms that Arp2/3 is required for model Hirano body formation in Dictyostelium.Fig. 5.


De novo actin polymerization is required for model Hirano body formation in Dictyostelium.

Dong Y, Shahid-Salles S, Sherling D, Fechheimer N, Iyer N, Wells L, Fechheimer M, Furukawa R - Biol Open (2016)

Model Hirano bodies form after removal of CK666. Cells were induced to express E60K-GFP and immediately incubated with CK666 for 12 h. They recovered in media after CK666 was removed for another 12 h (left column). Cells that were induced to express E60K-GFP at the same time (middle column) and at the time of CK666 removal (right column) were also observed as controls. All cells were examined by DIC (first row), and by fluorescence microscopy using TRITC-phalloidin (second row) or GFP (third row). Model Hirano bodies formed in a substantial amount of cells in all three samples. Scale bar=15 µm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4920178&req=5

BIO014944F5: Model Hirano bodies form after removal of CK666. Cells were induced to express E60K-GFP and immediately incubated with CK666 for 12 h. They recovered in media after CK666 was removed for another 12 h (left column). Cells that were induced to express E60K-GFP at the same time (middle column) and at the time of CK666 removal (right column) were also observed as controls. All cells were examined by DIC (first row), and by fluorescence microscopy using TRITC-phalloidin (second row) or GFP (third row). Model Hirano bodies formed in a substantial amount of cells in all three samples. Scale bar=15 µm.
Mentions: To rule out the possibility that the inhibition of model Hirano body formation by CK666 was due to a non-specific inhibition, we removed CK666 after 12 h of incubation and allowed the cells to recover for another 12 h with media under conditions for expression of E60K-GFP. Following removal of CK666, model Hirano bodies formed in a significant proportion of cells (Fig. 5). Thus, the effect of CK666 is reversible, and the process of model Hirano body formation can still be triggered. Therefore CK666, which inhibits the Arp2/3 complex activity, also inhibits model Hirano body formation and confirms that Arp2/3 is required for model Hirano body formation in Dictyostelium.Fig. 5.

Bottom Line: Whereas model Hirano bodies could form in WASH-deficient cells, they failed to form in cells lacking HSPC300, a member of the WAVE complex.In the case of VASP, both its G- and F-actin binding domains were required for model Hirano body formation.Collectively, our results indicate that de novo actin polymerization is required to form model Hirano bodies.

View Article: PubMed Central - PubMed

Affiliation: Department of Cellular Biology, University of Georgia, Athens, GA, USA 30602.

No MeSH data available.


Related in: MedlinePlus