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Regulatory mechanisms required for DE-cadherin function in cell migration and other types of adhesion.

Pacquelet A, Rørth P - J. Cell Biol. (2005)

Bottom Line: We have investigated the requirements for Drosophila melanogaster epithelial (DE) cadherin regulation in vivo.We found that (1) although linking DE-cadherin to alpha-catenin is essential, regulation of the link is not required in any of these types of adhesion; (2) beta-catenin is required only to link DE-cadherin to alpha-catenin; and (3) the cytoplasmic domain of DE-cadherin has an additional specific function for the invasive migration of border cells, which is conserved to other cadherins.The nature of this additional function is discussed.

View Article: PubMed Central - PubMed

Affiliation: European Molecular Biology Laboratory (EMBL), 69117 Heidelberg, Germany.

ABSTRACT
Cadherin-mediated adhesion can be regulated at many levels, as demonstrated by detailed analysis in cell lines. We have investigated the requirements for Drosophila melanogaster epithelial (DE) cadherin regulation in vivo. Investigating D. melanogaster oogenesis as a model system allowed the dissection of DE-cadherin function in several types of adhesion: cell sorting, cell positioning, epithelial integrity, and the cadherin-dependent process of border cell migration. We generated multiple fusions between DE-cadherin and alpha-catenin as well as point-mutated beta-catenin and analyzed their ability to support these types of adhesion. We found that (1) although linking DE-cadherin to alpha-catenin is essential, regulation of the link is not required in any of these types of adhesion; (2) beta-catenin is required only to link DE-cadherin to alpha-catenin; and (3) the cytoplasmic domain of DE-cadherin has an additional specific function for the invasive migration of border cells, which is conserved to other cadherins. The nature of this additional function is discussed.

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Activity of DE-cadherin with α-catenin in place of the cytoplasmic domain (DE-cadherinΔCyt/α-catenin). (A) Schematic representation of wild-type DE-cadherin and DE-cadherinΔCyt/α-catenin. (B–F′) Expression of wild-type DE-cadherin (B and D) and DE-cadherinΔCyt/α-catenin (C, E, and F) in shg- mutant (shgR69) follicle (B–E) and border cells (F). (B–E) Edge of the mutant clone (cells with increased staining) is indicated by the green line. (F) One border cell (asterisk) is mutant. (B, C, and F) All DE-cadherin is detected. (D and E) Only surface DE-cadherin is detected. (G) Oocyte positioning when follicle cells are shg- mutant and express the indicated transgenes. (H) Border cell migration in shg- mutant border cells expressing the indicated transgenes. Three independent transgenic lines were tested for DE-cadherinΔCyt/α-catenin. (I–J′) DE-cadherin levels in shgP34-1 mutant follicle (I) and border cell (J) clones. (I) Mutant cells are indicated by the presence of GFP (green). (J) Three border cells (asterisks) were mutant. Compare DE-cadherin staining between adjacent wild-type cells (arrowhead) with staining between adjacent mutant cells (arrow). (F, I, and J) Phalloidin (red) stains F-actin and DE-cadherin is in blue. Bars (B–E, I, and I′), 20 μm; (F, F′, J, and J′) 10 μm.
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fig3: Activity of DE-cadherin with α-catenin in place of the cytoplasmic domain (DE-cadherinΔCyt/α-catenin). (A) Schematic representation of wild-type DE-cadherin and DE-cadherinΔCyt/α-catenin. (B–F′) Expression of wild-type DE-cadherin (B and D) and DE-cadherinΔCyt/α-catenin (C, E, and F) in shg- mutant (shgR69) follicle (B–E) and border cells (F). (B–E) Edge of the mutant clone (cells with increased staining) is indicated by the green line. (F) One border cell (asterisk) is mutant. (B, C, and F) All DE-cadherin is detected. (D and E) Only surface DE-cadherin is detected. (G) Oocyte positioning when follicle cells are shg- mutant and express the indicated transgenes. (H) Border cell migration in shg- mutant border cells expressing the indicated transgenes. Three independent transgenic lines were tested for DE-cadherinΔCyt/α-catenin. (I–J′) DE-cadherin levels in shgP34-1 mutant follicle (I) and border cell (J) clones. (I) Mutant cells are indicated by the presence of GFP (green). (J) Three border cells (asterisks) were mutant. Compare DE-cadherin staining between adjacent wild-type cells (arrowhead) with staining between adjacent mutant cells (arrow). (F, I, and J) Phalloidin (red) stains F-actin and DE-cadherin is in blue. Bars (B–E, I, and I′), 20 μm; (F, F′, J, and J′) 10 μm.

Mentions: DE-cadherinΔCyt/α-catenin was obtained by fusing full-length (FL) α-catenin directly after the transmembrane domain of DE-cadherin (Fig. 3 A). This fusion protein should link DE-cadherin to the cytoskeleton but prevent any regulation of the link between DE-cadherin and β-catenin as well as between β- and α-catenin. Functionality in vivo was analyzed by the ability to provide DE-cadherin function to cells lacking endogenous shg (DE-cadherin). Both wild-type DE-cadherin and DE-cadherinΔCyt/α-catenin–expressing transgenes rescued the phenotypes that were observed in shg mutant follicle cell clones (follicle cell sorting, loss of epithelial integrity, and oocyte mispositioning; Fig. 3, E and G). Thus, DE-cadherinΔCyt/α-catenin can substitute for endogenous DE-cadherin in follicle cells. This shows that it is able to mediate productive adhesion. Wild-type DE-cadherin fully rescued migration defects of shg mutant border cells (Fig. 3 H). In contrast, DE-cadherinΔCyt/α-catenin showed almost no rescue ability in border cells (Fig. 3 H).


Regulatory mechanisms required for DE-cadherin function in cell migration and other types of adhesion.

Pacquelet A, Rørth P - J. Cell Biol. (2005)

Activity of DE-cadherin with α-catenin in place of the cytoplasmic domain (DE-cadherinΔCyt/α-catenin). (A) Schematic representation of wild-type DE-cadherin and DE-cadherinΔCyt/α-catenin. (B–F′) Expression of wild-type DE-cadherin (B and D) and DE-cadherinΔCyt/α-catenin (C, E, and F) in shg- mutant (shgR69) follicle (B–E) and border cells (F). (B–E) Edge of the mutant clone (cells with increased staining) is indicated by the green line. (F) One border cell (asterisk) is mutant. (B, C, and F) All DE-cadherin is detected. (D and E) Only surface DE-cadherin is detected. (G) Oocyte positioning when follicle cells are shg- mutant and express the indicated transgenes. (H) Border cell migration in shg- mutant border cells expressing the indicated transgenes. Three independent transgenic lines were tested for DE-cadherinΔCyt/α-catenin. (I–J′) DE-cadherin levels in shgP34-1 mutant follicle (I) and border cell (J) clones. (I) Mutant cells are indicated by the presence of GFP (green). (J) Three border cells (asterisks) were mutant. Compare DE-cadherin staining between adjacent wild-type cells (arrowhead) with staining between adjacent mutant cells (arrow). (F, I, and J) Phalloidin (red) stains F-actin and DE-cadherin is in blue. Bars (B–E, I, and I′), 20 μm; (F, F′, J, and J′) 10 μm.
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getmorefigures.php?uid=PMC2171345&req=5

fig3: Activity of DE-cadherin with α-catenin in place of the cytoplasmic domain (DE-cadherinΔCyt/α-catenin). (A) Schematic representation of wild-type DE-cadherin and DE-cadherinΔCyt/α-catenin. (B–F′) Expression of wild-type DE-cadherin (B and D) and DE-cadherinΔCyt/α-catenin (C, E, and F) in shg- mutant (shgR69) follicle (B–E) and border cells (F). (B–E) Edge of the mutant clone (cells with increased staining) is indicated by the green line. (F) One border cell (asterisk) is mutant. (B, C, and F) All DE-cadherin is detected. (D and E) Only surface DE-cadherin is detected. (G) Oocyte positioning when follicle cells are shg- mutant and express the indicated transgenes. (H) Border cell migration in shg- mutant border cells expressing the indicated transgenes. Three independent transgenic lines were tested for DE-cadherinΔCyt/α-catenin. (I–J′) DE-cadherin levels in shgP34-1 mutant follicle (I) and border cell (J) clones. (I) Mutant cells are indicated by the presence of GFP (green). (J) Three border cells (asterisks) were mutant. Compare DE-cadherin staining between adjacent wild-type cells (arrowhead) with staining between adjacent mutant cells (arrow). (F, I, and J) Phalloidin (red) stains F-actin and DE-cadherin is in blue. Bars (B–E, I, and I′), 20 μm; (F, F′, J, and J′) 10 μm.
Mentions: DE-cadherinΔCyt/α-catenin was obtained by fusing full-length (FL) α-catenin directly after the transmembrane domain of DE-cadherin (Fig. 3 A). This fusion protein should link DE-cadherin to the cytoskeleton but prevent any regulation of the link between DE-cadherin and β-catenin as well as between β- and α-catenin. Functionality in vivo was analyzed by the ability to provide DE-cadherin function to cells lacking endogenous shg (DE-cadherin). Both wild-type DE-cadherin and DE-cadherinΔCyt/α-catenin–expressing transgenes rescued the phenotypes that were observed in shg mutant follicle cell clones (follicle cell sorting, loss of epithelial integrity, and oocyte mispositioning; Fig. 3, E and G). Thus, DE-cadherinΔCyt/α-catenin can substitute for endogenous DE-cadherin in follicle cells. This shows that it is able to mediate productive adhesion. Wild-type DE-cadherin fully rescued migration defects of shg mutant border cells (Fig. 3 H). In contrast, DE-cadherinΔCyt/α-catenin showed almost no rescue ability in border cells (Fig. 3 H).

Bottom Line: We have investigated the requirements for Drosophila melanogaster epithelial (DE) cadherin regulation in vivo.We found that (1) although linking DE-cadherin to alpha-catenin is essential, regulation of the link is not required in any of these types of adhesion; (2) beta-catenin is required only to link DE-cadherin to alpha-catenin; and (3) the cytoplasmic domain of DE-cadherin has an additional specific function for the invasive migration of border cells, which is conserved to other cadherins.The nature of this additional function is discussed.

View Article: PubMed Central - PubMed

Affiliation: European Molecular Biology Laboratory (EMBL), 69117 Heidelberg, Germany.

ABSTRACT
Cadherin-mediated adhesion can be regulated at many levels, as demonstrated by detailed analysis in cell lines. We have investigated the requirements for Drosophila melanogaster epithelial (DE) cadherin regulation in vivo. Investigating D. melanogaster oogenesis as a model system allowed the dissection of DE-cadherin function in several types of adhesion: cell sorting, cell positioning, epithelial integrity, and the cadherin-dependent process of border cell migration. We generated multiple fusions between DE-cadherin and alpha-catenin as well as point-mutated beta-catenin and analyzed their ability to support these types of adhesion. We found that (1) although linking DE-cadherin to alpha-catenin is essential, regulation of the link is not required in any of these types of adhesion; (2) beta-catenin is required only to link DE-cadherin to alpha-catenin; and (3) the cytoplasmic domain of DE-cadherin has an additional specific function for the invasive migration of border cells, which is conserved to other cadherins. The nature of this additional function is discussed.

Show MeSH
Related in: MedlinePlus