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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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Confocal immunofluorescence microscopy of MDCK  cells transiently transfected with  δ-catenin. The cells were double  labeled with (A) δ-catenin antibodies and with (B) desmoplakin antibody. The arrow  points to the transfected cell. (C)  Merged fluorescent image showing minimal colocalization of δ-catenin and desmoplakin. The horizontal line indicates where the  XZ plane was selected for D–F.  (D–F) Respective XZ vertical  sections of A–C. Bar, 15 μm.
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Figure 5: Confocal immunofluorescence microscopy of MDCK cells transiently transfected with δ-catenin. The cells were double labeled with (A) δ-catenin antibodies and with (B) desmoplakin antibody. The arrow points to the transfected cell. (C) Merged fluorescent image showing minimal colocalization of δ-catenin and desmoplakin. The horizontal line indicates where the XZ plane was selected for D–F. (D–F) Respective XZ vertical sections of A–C. Bar, 15 μm.

Mentions: Because δ-catenin shares significant homology with p0071, a desmosomal protein (Hatzfeld and Nachtsheim, 1996), we compared the localization of δ-catenin with both desmoplakin and E-cadherin (Figs. 4 and 5). MDCK cells were transiently transfected with δ-catenin and double labeled with δ-catenin antibodies and either desmoplakin or E-cadherin antibodies. In transfected cells, δ-catenin codistributed with the adherens junction protein, E-cadherin in a thick band around the cell periphery, while neighboring untransfected cells retained a typical less intense junctional pattern of immunoreactivity (Fig. 4, A–C). Double labeling immunofluorescent microscopy showed that δ-catenin even colocalized with E-cadherin along cell processes (Fig. 4, A–C). Because δ-catenin–transfected cells are less flat than nontransfected MDCK cells, a z-series showed that δ-catenin and E-cadherin also codistributed in the vertical axis (Fig. 4, D–F). The greater intensity of E-cadherin staining in δ-catenin–transfected cells suggested that the concentration of E-cadherin in the transfected cells increased (Fig. 4, A and B). Densitometric measurement of protein profiles on immunoblots demonstrated a 30% increase in the E-cadherin level while p120ctn and desmoglein levels did not significantly change (data not shown). Double labeling immunofluorescence with δ-catenin and desmoplakin revealed a less close relationship (Fig. 5, A–F). In δ-catenin–transfected cells, the intensity of the desmoplakin immunoreactivity did not increase, and the pattern of desmoplakin labeling did not completely codistribute with δ-catenin. Desmoplakin labeling did not extend into cell processes (Fig. 5, A–C) and did not extend apically in the z-axis (Fig. 5, D–F). These findings do not exclude any colocalization between desmoplakin and δ-catenin, but they clearly point to a preferential cellular relationship to E-cadherin. δ-catenin also did not colocalize with the tight-junction associated protein, ZO-1 (data not shown). We therefore concluded that exogenous δ-catenin can be directed to the adherens junctions in MDCK cells.


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)

Confocal immunofluorescence microscopy of MDCK  cells transiently transfected with  δ-catenin. The cells were double  labeled with (A) δ-catenin antibodies and with (B) desmoplakin antibody. The arrow  points to the transfected cell. (C)  Merged fluorescent image showing minimal colocalization of δ-catenin and desmoplakin. The horizontal line indicates where the  XZ plane was selected for D–F.  (D–F) Respective XZ vertical  sections of A–C. Bar, 15 μm.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2132907&req=5

Figure 5: Confocal immunofluorescence microscopy of MDCK cells transiently transfected with δ-catenin. The cells were double labeled with (A) δ-catenin antibodies and with (B) desmoplakin antibody. The arrow points to the transfected cell. (C) Merged fluorescent image showing minimal colocalization of δ-catenin and desmoplakin. The horizontal line indicates where the XZ plane was selected for D–F. (D–F) Respective XZ vertical sections of A–C. Bar, 15 μm.
Mentions: Because δ-catenin shares significant homology with p0071, a desmosomal protein (Hatzfeld and Nachtsheim, 1996), we compared the localization of δ-catenin with both desmoplakin and E-cadherin (Figs. 4 and 5). MDCK cells were transiently transfected with δ-catenin and double labeled with δ-catenin antibodies and either desmoplakin or E-cadherin antibodies. In transfected cells, δ-catenin codistributed with the adherens junction protein, E-cadherin in a thick band around the cell periphery, while neighboring untransfected cells retained a typical less intense junctional pattern of immunoreactivity (Fig. 4, A–C). Double labeling immunofluorescent microscopy showed that δ-catenin even colocalized with E-cadherin along cell processes (Fig. 4, A–C). Because δ-catenin–transfected cells are less flat than nontransfected MDCK cells, a z-series showed that δ-catenin and E-cadherin also codistributed in the vertical axis (Fig. 4, D–F). The greater intensity of E-cadherin staining in δ-catenin–transfected cells suggested that the concentration of E-cadherin in the transfected cells increased (Fig. 4, A and B). Densitometric measurement of protein profiles on immunoblots demonstrated a 30% increase in the E-cadherin level while p120ctn and desmoglein levels did not significantly change (data not shown). Double labeling immunofluorescence with δ-catenin and desmoplakin revealed a less close relationship (Fig. 5, A–F). In δ-catenin–transfected cells, the intensity of the desmoplakin immunoreactivity did not increase, and the pattern of desmoplakin labeling did not completely codistribute with δ-catenin. Desmoplakin labeling did not extend into cell processes (Fig. 5, A–C) and did not extend apically in the z-axis (Fig. 5, D–F). These findings do not exclude any colocalization between desmoplakin and δ-catenin, but they clearly point to a preferential cellular relationship to E-cadherin. δ-catenin also did not colocalize with the tight-junction associated protein, ZO-1 (data not shown). We therefore concluded that exogenous δ-catenin can be directed to the adherens junctions in MDCK cells.

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