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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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Expression of δ-catenin cDNA in transfected MDCK  cells and endogenous δ-catenin  in developing mouse brains. (A)  Immunoblot showing δ-catenin  expression in MDCK cells and  developing mouse brains. (1)  Mock-transfected MDCK cells;  (2) MF (MDCK cells transfected with full-length δ-catenin  cDNA); (3) MB (mouse brain lysate). (B) Immunoblot of transfected MDCK cells showing  δ-catenin in the 0.2% Triton  X-100 soluble (TXs, lane 1) and  insoluble (Txi, lane 2) fractions.  (C) Immunoblots of mock (1 and  2) and δ-catenin transfected (3 and 4) MDCK cells extracted in 1.0% Triton X-100. Soluble and insoluble fractions were blotted with  the indicated antibodies. In all panels the molecular weight standard is indicated at the left.
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Figure 2: Expression of δ-catenin cDNA in transfected MDCK cells and endogenous δ-catenin in developing mouse brains. (A) Immunoblot showing δ-catenin expression in MDCK cells and developing mouse brains. (1) Mock-transfected MDCK cells; (2) MF (MDCK cells transfected with full-length δ-catenin cDNA); (3) MB (mouse brain lysate). (B) Immunoblot of transfected MDCK cells showing δ-catenin in the 0.2% Triton X-100 soluble (TXs, lane 1) and insoluble (Txi, lane 2) fractions. (C) Immunoblots of mock (1 and 2) and δ-catenin transfected (3 and 4) MDCK cells extracted in 1.0% Triton X-100. Soluble and insoluble fractions were blotted with the indicated antibodies. In all panels the molecular weight standard is indicated at the left.

Mentions: We raised two antibodies directed against different δ-catenin epitopes. Both specifically recognized a single band at 160 kD in MDCK cells transiently transfected with a full-length δ-catenin cDNA, but did not react with mock-transfected cells (Fig. 2 A). Because δ-catenin is specifically expressed in the nervous system (Zhou et al., 1997), postnatal day 2 mouse brain was also used for an immunoblot analysis. In this assay, δ-catenin migrated as a doublet at 160 kD (Fig. 2 A). The retarded mobility of δ-catenin relative to its predicted molecular weight may be due to the presence of charge groups, posttranslational modifications, or an extended structure which affects its electrophoretic migration.


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)

Expression of δ-catenin cDNA in transfected MDCK  cells and endogenous δ-catenin  in developing mouse brains. (A)  Immunoblot showing δ-catenin  expression in MDCK cells and  developing mouse brains. (1)  Mock-transfected MDCK cells;  (2) MF (MDCK cells transfected with full-length δ-catenin  cDNA); (3) MB (mouse brain lysate). (B) Immunoblot of transfected MDCK cells showing  δ-catenin in the 0.2% Triton  X-100 soluble (TXs, lane 1) and  insoluble (Txi, lane 2) fractions.  (C) Immunoblots of mock (1 and  2) and δ-catenin transfected (3 and 4) MDCK cells extracted in 1.0% Triton X-100. Soluble and insoluble fractions were blotted with  the indicated antibodies. In all panels the molecular weight standard is indicated at the left.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2132907&req=5

Figure 2: Expression of δ-catenin cDNA in transfected MDCK cells and endogenous δ-catenin in developing mouse brains. (A) Immunoblot showing δ-catenin expression in MDCK cells and developing mouse brains. (1) Mock-transfected MDCK cells; (2) MF (MDCK cells transfected with full-length δ-catenin cDNA); (3) MB (mouse brain lysate). (B) Immunoblot of transfected MDCK cells showing δ-catenin in the 0.2% Triton X-100 soluble (TXs, lane 1) and insoluble (Txi, lane 2) fractions. (C) Immunoblots of mock (1 and 2) and δ-catenin transfected (3 and 4) MDCK cells extracted in 1.0% Triton X-100. Soluble and insoluble fractions were blotted with the indicated antibodies. In all panels the molecular weight standard is indicated at the left.
Mentions: We raised two antibodies directed against different δ-catenin epitopes. Both specifically recognized a single band at 160 kD in MDCK cells transiently transfected with a full-length δ-catenin cDNA, but did not react with mock-transfected cells (Fig. 2 A). Because δ-catenin is specifically expressed in the nervous system (Zhou et al., 1997), postnatal day 2 mouse brain was also used for an immunoblot analysis. In this assay, δ-catenin migrated as a doublet at 160 kD (Fig. 2 A). The retarded mobility of δ-catenin relative to its predicted molecular weight may be due to the presence of charge groups, posttranslational modifications, or an extended structure which affects its electrophoretic migration.

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