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delta-catenin, an adhesive junction-associated protein which promotes cell scattering.

Lu Q, Paredes M, Medina M, Zhou J, Cavallo R, Peifer M, Orecchio L, Kosik KS - J. Cell Biol. (1999)

Bottom Line: We found that delta-catenin can be immunoprecipitated as a complex with other components of the adherens junction, including cadherin and beta-catenin, from transfected cells and brain.In developing mouse brain, staining with delta-catenin antibodies is prominent towards the apical boundary of the neuroepithelial cells in the ventricular zone.The Arm domain alone was sufficient for achieving localization and coimmunoprecipitation with cadherin.

View Article: PubMed Central - PubMed

Affiliation: Center for Neurologic Diseases, Department of Neurology, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts 02115, USA.

ABSTRACT
The classical adherens junction that holds epithelial cells together consists of a protein complex in which members of the cadherin family linked to various catenins are the principal components. delta-catenin is a mammalian brain protein in the Armadillo repeat superfamily with sequence similarity to the adherens junction protein p120(ctn). We found that delta-catenin can be immunoprecipitated as a complex with other components of the adherens junction, including cadherin and beta-catenin, from transfected cells and brain. The interaction with cadherin involves direct contact within the highly conserved juxtamembrane region of the COOH terminus, where p120(ctn) also binds. In developing mouse brain, staining with delta-catenin antibodies is prominent towards the apical boundary of the neuroepithelial cells in the ventricular zone. When transfected into Madin-Darby canine kidney (MDCK) epithelial cells delta-catenin colocalized with cadherin, p120(ctn), and beta-catenin. The Arm domain alone was sufficient for achieving localization and coimmunoprecipitation with cadherin. The ectopic expression of delta-catenin in MDCK cells altered their morphology, induced the elaboration of lamellipodia, interfered with monolayer formation, and increased scattering in response to hepatocyte growth factor treatment. We propose that delta-catenin can regulate adhesion molecules to implement the organization of large cellular arrays necessary for tissue morphogenesis.

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Association of δ-catenin with adhesive junction proteins in brain and in MDCK cells  stably expressing δ-catenin. (A)  δ-catenin coimmunoprecipitates  E-cadherin and β-catenin, but  not desmoglein. (1 and 6) Mock-transfected MDCK cells. (2–5, 8,  and 9) MF cells. (B) Reverse  immunoprecipitation showing  δ-catenin coprecipitated with  E-cadherin and β-catenin. (1)  Mock-transfected MDCK cells.  (2–5) MF cells. (C) δ-catenin  coimmunoprecipitates N-cadherin and β-catenin in brains. (1)  N-cadherin immunoblot of brain  fractions immunoprecipitated  using nonimmune rabbit IgG.  (2) N-cadherin immunoblot of  brain fractions immunoprecipitated using rAb62. (3) N-cadherin immunoblot of brain lysate. (4) β-catenin immunoblot  of brain fractions immunoprecipitated using rAb62. (D–F)  Cofractionation of brain δ-catenin with N-cadherin and synaptophysin. PnH, postnuclear  homogenates. P2, heavy membranes consisting of myelin, mitochondria, and crude synaptosomes. P3, mostly microsomes. SPM, synaptic plasma membranes. (D) Immunoblot showing brain fractionation profile of synaptophysin. (E) Immunoblot showing brain fractionation profile of δ-catenin. (F)  Immunoblot showing brain fractionation profile of N-cadherin. Molecular weight markers are indicated at the left of each panel.
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Figure 6: Association of δ-catenin with adhesive junction proteins in brain and in MDCK cells stably expressing δ-catenin. (A) δ-catenin coimmunoprecipitates E-cadherin and β-catenin, but not desmoglein. (1 and 6) Mock-transfected MDCK cells. (2–5, 8, and 9) MF cells. (B) Reverse immunoprecipitation showing δ-catenin coprecipitated with E-cadherin and β-catenin. (1) Mock-transfected MDCK cells. (2–5) MF cells. (C) δ-catenin coimmunoprecipitates N-cadherin and β-catenin in brains. (1) N-cadherin immunoblot of brain fractions immunoprecipitated using nonimmune rabbit IgG. (2) N-cadherin immunoblot of brain fractions immunoprecipitated using rAb62. (3) N-cadherin immunoblot of brain lysate. (4) β-catenin immunoblot of brain fractions immunoprecipitated using rAb62. (D–F) Cofractionation of brain δ-catenin with N-cadherin and synaptophysin. PnH, postnuclear homogenates. P2, heavy membranes consisting of myelin, mitochondria, and crude synaptosomes. P3, mostly microsomes. SPM, synaptic plasma membranes. (D) Immunoblot showing brain fractionation profile of synaptophysin. (E) Immunoblot showing brain fractionation profile of δ-catenin. (F) Immunoblot showing brain fractionation profile of N-cadherin. Molecular weight markers are indicated at the left of each panel.

Mentions: Based on colocalization data we asked whether δ-catenin interacts with the adherens junction proteins E-cadherin and β-catenin. In MDCK cells stably expressing δ-catenin, coimmunoprecipitation experiments indicated that δ-catenin coimmunoprecipitated both E-cadherin, β-catenin (Fig. 6 A), and p120ctn efficiently (not shown), but did not bring down desmoglein, which is abundantly present in MDCK cells and reacted with the desmoglein antibody (Fig. 6 A, lane 9). We concluded that at most, only minimal amounts of δ-catenin are associated with desmosomes. In reverse immunoprecipitation experiments, both β-catenin and E-cadherin coprecipitated δ-catenin (Fig. 6 B, lanes 2 and 3). The interaction between δ-catenin and E-cadherin/β-catenin was abolished in RIPA buffer that contained 0.2% SDS, although the coimmunoprecipitation of E-cadherin with β-catenin was retained (data not shown).


delta-catenin, an adhesive junction-associated protein which promotes cell scattering.

Lu Q, Paredes M, Medina M, Zhou J, Cavallo R, Peifer M, Orecchio L, Kosik KS - J. Cell Biol. (1999)

Association of δ-catenin with adhesive junction proteins in brain and in MDCK cells  stably expressing δ-catenin. (A)  δ-catenin coimmunoprecipitates  E-cadherin and β-catenin, but  not desmoglein. (1 and 6) Mock-transfected MDCK cells. (2–5, 8,  and 9) MF cells. (B) Reverse  immunoprecipitation showing  δ-catenin coprecipitated with  E-cadherin and β-catenin. (1)  Mock-transfected MDCK cells.  (2–5) MF cells. (C) δ-catenin  coimmunoprecipitates N-cadherin and β-catenin in brains. (1)  N-cadherin immunoblot of brain  fractions immunoprecipitated  using nonimmune rabbit IgG.  (2) N-cadherin immunoblot of  brain fractions immunoprecipitated using rAb62. (3) N-cadherin immunoblot of brain lysate. (4) β-catenin immunoblot  of brain fractions immunoprecipitated using rAb62. (D–F)  Cofractionation of brain δ-catenin with N-cadherin and synaptophysin. PnH, postnuclear  homogenates. P2, heavy membranes consisting of myelin, mitochondria, and crude synaptosomes. P3, mostly microsomes. SPM, synaptic plasma membranes. (D) Immunoblot showing brain fractionation profile of synaptophysin. (E) Immunoblot showing brain fractionation profile of δ-catenin. (F)  Immunoblot showing brain fractionation profile of N-cadherin. Molecular weight markers are indicated at the left of each panel.
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Figure 6: Association of δ-catenin with adhesive junction proteins in brain and in MDCK cells stably expressing δ-catenin. (A) δ-catenin coimmunoprecipitates E-cadherin and β-catenin, but not desmoglein. (1 and 6) Mock-transfected MDCK cells. (2–5, 8, and 9) MF cells. (B) Reverse immunoprecipitation showing δ-catenin coprecipitated with E-cadherin and β-catenin. (1) Mock-transfected MDCK cells. (2–5) MF cells. (C) δ-catenin coimmunoprecipitates N-cadherin and β-catenin in brains. (1) N-cadherin immunoblot of brain fractions immunoprecipitated using nonimmune rabbit IgG. (2) N-cadherin immunoblot of brain fractions immunoprecipitated using rAb62. (3) N-cadherin immunoblot of brain lysate. (4) β-catenin immunoblot of brain fractions immunoprecipitated using rAb62. (D–F) Cofractionation of brain δ-catenin with N-cadherin and synaptophysin. PnH, postnuclear homogenates. P2, heavy membranes consisting of myelin, mitochondria, and crude synaptosomes. P3, mostly microsomes. SPM, synaptic plasma membranes. (D) Immunoblot showing brain fractionation profile of synaptophysin. (E) Immunoblot showing brain fractionation profile of δ-catenin. (F) Immunoblot showing brain fractionation profile of N-cadherin. Molecular weight markers are indicated at the left of each panel.
Mentions: Based on colocalization data we asked whether δ-catenin interacts with the adherens junction proteins E-cadherin and β-catenin. In MDCK cells stably expressing δ-catenin, coimmunoprecipitation experiments indicated that δ-catenin coimmunoprecipitated both E-cadherin, β-catenin (Fig. 6 A), and p120ctn efficiently (not shown), but did not bring down desmoglein, which is abundantly present in MDCK cells and reacted with the desmoglein antibody (Fig. 6 A, lane 9). We concluded that at most, only minimal amounts of δ-catenin are associated with desmosomes. In reverse immunoprecipitation experiments, both β-catenin and E-cadherin coprecipitated δ-catenin (Fig. 6 B, lanes 2 and 3). The interaction between δ-catenin and E-cadherin/β-catenin was abolished in RIPA buffer that contained 0.2% SDS, although the coimmunoprecipitation of E-cadherin with β-catenin was retained (data not shown).

Bottom Line: We found that delta-catenin can be immunoprecipitated as a complex with other components of the adherens junction, including cadherin and beta-catenin, from transfected cells and brain.In developing mouse brain, staining with delta-catenin antibodies is prominent towards the apical boundary of the neuroepithelial cells in the ventricular zone.The Arm domain alone was sufficient for achieving localization and coimmunoprecipitation with cadherin.

View Article: PubMed Central - PubMed

Affiliation: Center for Neurologic Diseases, Department of Neurology, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts 02115, USA.

ABSTRACT
The classical adherens junction that holds epithelial cells together consists of a protein complex in which members of the cadherin family linked to various catenins are the principal components. delta-catenin is a mammalian brain protein in the Armadillo repeat superfamily with sequence similarity to the adherens junction protein p120(ctn). We found that delta-catenin can be immunoprecipitated as a complex with other components of the adherens junction, including cadherin and beta-catenin, from transfected cells and brain. The interaction with cadherin involves direct contact within the highly conserved juxtamembrane region of the COOH terminus, where p120(ctn) also binds. In developing mouse brain, staining with delta-catenin antibodies is prominent towards the apical boundary of the neuroepithelial cells in the ventricular zone. When transfected into Madin-Darby canine kidney (MDCK) epithelial cells delta-catenin colocalized with cadherin, p120(ctn), and beta-catenin. The Arm domain alone was sufficient for achieving localization and coimmunoprecipitation with cadherin. The ectopic expression of delta-catenin in MDCK cells altered their morphology, induced the elaboration of lamellipodia, interfered with monolayer formation, and increased scattering in response to hepatocyte growth factor treatment. We propose that delta-catenin can regulate adhesion molecules to implement the organization of large cellular arrays necessary for tissue morphogenesis.

Show MeSH
Related in: MedlinePlus