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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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Deletion analysis of  δ-catenin localization and interaction with adhesive junction  proteins. (A) Schematic drawings show the design of deletion  mutants. MF, full-length δ-catenin. ARM1-10, δ-catenin sequence containing only the  Arm repeats. ARM/NLS,  δ-catenin containing partial  NH2 terminus including six  Arm repeats. Within Arm repeat 6, the boxed sequence indicates a putative nuclear localization signal. ARM/CT212:  δ-catenin containing the complete COOH terminus but with  only four Arm repeats from the  COOH-terminal end. (B) Localization of mutant δ-catenin  in MDCK cells. (a) Mock transfection. (b) GFP reporter. (c)  Full-length δ-catenin. (d) Armadillo domain alone. (e)  ARM/NLS. (f) ARM/CT212.  Bar, 10 μm. (C) Coimmunoprecipitation of δ-catenin deletion  mutants from transfected  MDCK cells. Fractions were  immunoprecipitated with GFP  antibody and labeled with either E-cadherin or β-catenin antibody. (1) Mock-transfected cells. (2) MF. (3) ARM1-10. (4) ARM/ CT212. (5) ARM/NLS. Molecular weight markers are indicated at the left of each panel.
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Figure 8: Deletion analysis of δ-catenin localization and interaction with adhesive junction proteins. (A) Schematic drawings show the design of deletion mutants. MF, full-length δ-catenin. ARM1-10, δ-catenin sequence containing only the Arm repeats. ARM/NLS, δ-catenin containing partial NH2 terminus including six Arm repeats. Within Arm repeat 6, the boxed sequence indicates a putative nuclear localization signal. ARM/CT212: δ-catenin containing the complete COOH terminus but with only four Arm repeats from the COOH-terminal end. (B) Localization of mutant δ-catenin in MDCK cells. (a) Mock transfection. (b) GFP reporter. (c) Full-length δ-catenin. (d) Armadillo domain alone. (e) ARM/NLS. (f) ARM/CT212. Bar, 10 μm. (C) Coimmunoprecipitation of δ-catenin deletion mutants from transfected MDCK cells. Fractions were immunoprecipitated with GFP antibody and labeled with either E-cadherin or β-catenin antibody. (1) Mock-transfected cells. (2) MF. (3) ARM1-10. (4) ARM/ CT212. (5) ARM/NLS. Molecular weight markers are indicated at the left of each panel.

Mentions: We generated a number of δ-catenin deletion mutants to define the domains necessary for targeting δ-catenin to the adherens junction (Fig. 8 A). To visualize these deletion mutants directly following transfection, full-length and various mutant constructs were fused with green fluorescent protein (GFP) and subcloned into the pEGFP vector. Full-length δ-catenin fused to EGFP showed a fluorescence distribution pattern that was identical to the antibody-labeling pattern of cells transfected with δ-catenin alone (Fig. 8 B, panel c). Although the primary site of δ-catenin immunoreactivity is the cell–cell junction, immunolabeling of cells also suggested a cytoplasmic pool. When both the NH2- and COOH-terminal extensions were deleted, leaving only the arm repeats, δ-catenin maintained its localization to cell–cell junctions based on detection of the GFP marker (Fig. 8 B, panel d) and on coimmunoprecipitation with E-cadherin and β-catenin (Fig. 8 C, lane 3). A significantly reduced interaction was observed for those constructs with truncations in the Arm repeat region (Fig. 8 C, lanes 4 and 5). Thus the armadillo repeat domain alone was sufficient for δ-catenin localization to cell–cell junctions as is the case for β-catenin which requires only a portion of the Arm domain to bind cadherin and localize to adherens junctions (Funayama et al., 1995; Orsulic and Peifer, 1996). These data are consistent with the two-hybrid analysis described above which also demonstrated that the δ-catenin Arm domain was sufficient for interacting directly with cadherin.


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)

Deletion analysis of  δ-catenin localization and interaction with adhesive junction  proteins. (A) Schematic drawings show the design of deletion  mutants. MF, full-length δ-catenin. ARM1-10, δ-catenin sequence containing only the  Arm repeats. ARM/NLS,  δ-catenin containing partial  NH2 terminus including six  Arm repeats. Within Arm repeat 6, the boxed sequence indicates a putative nuclear localization signal. ARM/CT212:  δ-catenin containing the complete COOH terminus but with  only four Arm repeats from the  COOH-terminal end. (B) Localization of mutant δ-catenin  in MDCK cells. (a) Mock transfection. (b) GFP reporter. (c)  Full-length δ-catenin. (d) Armadillo domain alone. (e)  ARM/NLS. (f) ARM/CT212.  Bar, 10 μm. (C) Coimmunoprecipitation of δ-catenin deletion  mutants from transfected  MDCK cells. Fractions were  immunoprecipitated with GFP  antibody and labeled with either E-cadherin or β-catenin antibody. (1) Mock-transfected cells. (2) MF. (3) ARM1-10. (4) ARM/ CT212. (5) ARM/NLS. Molecular weight markers are indicated at the left of each panel.
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Figure 8: Deletion analysis of δ-catenin localization and interaction with adhesive junction proteins. (A) Schematic drawings show the design of deletion mutants. MF, full-length δ-catenin. ARM1-10, δ-catenin sequence containing only the Arm repeats. ARM/NLS, δ-catenin containing partial NH2 terminus including six Arm repeats. Within Arm repeat 6, the boxed sequence indicates a putative nuclear localization signal. ARM/CT212: δ-catenin containing the complete COOH terminus but with only four Arm repeats from the COOH-terminal end. (B) Localization of mutant δ-catenin in MDCK cells. (a) Mock transfection. (b) GFP reporter. (c) Full-length δ-catenin. (d) Armadillo domain alone. (e) ARM/NLS. (f) ARM/CT212. Bar, 10 μm. (C) Coimmunoprecipitation of δ-catenin deletion mutants from transfected MDCK cells. Fractions were immunoprecipitated with GFP antibody and labeled with either E-cadherin or β-catenin antibody. (1) Mock-transfected cells. (2) MF. (3) ARM1-10. (4) ARM/ CT212. (5) ARM/NLS. Molecular weight markers are indicated at the left of each panel.
Mentions: We generated a number of δ-catenin deletion mutants to define the domains necessary for targeting δ-catenin to the adherens junction (Fig. 8 A). To visualize these deletion mutants directly following transfection, full-length and various mutant constructs were fused with green fluorescent protein (GFP) and subcloned into the pEGFP vector. Full-length δ-catenin fused to EGFP showed a fluorescence distribution pattern that was identical to the antibody-labeling pattern of cells transfected with δ-catenin alone (Fig. 8 B, panel c). Although the primary site of δ-catenin immunoreactivity is the cell–cell junction, immunolabeling of cells also suggested a cytoplasmic pool. When both the NH2- and COOH-terminal extensions were deleted, leaving only the arm repeats, δ-catenin maintained its localization to cell–cell junctions based on detection of the GFP marker (Fig. 8 B, panel d) and on coimmunoprecipitation with E-cadherin and β-catenin (Fig. 8 C, lane 3). A significantly reduced interaction was observed for those constructs with truncations in the Arm repeat region (Fig. 8 C, lanes 4 and 5). Thus the armadillo repeat domain alone was sufficient for δ-catenin localization to cell–cell junctions as is the case for β-catenin which requires only a portion of the Arm domain to bind cadherin and localize to adherens junctions (Funayama et al., 1995; Orsulic and Peifer, 1996). These data are consistent with the two-hybrid analysis described above which also demonstrated that the δ-catenin Arm domain was sufficient for interacting directly with cadherin.

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