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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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δ-catenin interacts directly with the juxtamembrane region of cadherins in the yeast two-hybrid system. (A) Interaction of  δ-catenin with various cadherin fragments. The Arm repeat region of human δ-catenin in the pCK2 “bait” vector was tested against  pCK4 vector alone or with pCK4 fusions encoding the full-length murine E-cadherin cytoplasmic domain (pCK4 ME-CAD cyto), two  complementary fragments of murine E-cadherin carrying either the membrane-proximal region (pCK4 MEC2) or the distal region with  the β-catenin binding site (pCK4 MEC3), a COOH-terminal fragment of OB-cadherin not containing the juxtamembrane region  (pCK4 OB-CAD CT), the entire cytoplasmic domain of Drosophila E-cadherin (pCK4 DEC), and smaller fragments of Drosophila  E-cadherin as shown in B (pCK4 DEC 4, 14, and 15). (B) Schematic summary of the cadherin fragments and their interaction with  δ-catenin. (C) Sequence alignment of the cytoplasmic tails of mouse E-cadherin, mouse OB-cadherin, and Drosophila DE-cadherin.  Above the sequences are shown the smallest fragment of mouse E-cadherin which bound δ-catenin in the yeast two-hybrid assay, while  below are diagrammed the amino acids missing from the OB-cadherin clone which does not bind to δ-catenin. The β-catenin/Armadillo  binding site is also indicated.
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Figure 7: δ-catenin interacts directly with the juxtamembrane region of cadherins in the yeast two-hybrid system. (A) Interaction of δ-catenin with various cadherin fragments. The Arm repeat region of human δ-catenin in the pCK2 “bait” vector was tested against pCK4 vector alone or with pCK4 fusions encoding the full-length murine E-cadherin cytoplasmic domain (pCK4 ME-CAD cyto), two complementary fragments of murine E-cadherin carrying either the membrane-proximal region (pCK4 MEC2) or the distal region with the β-catenin binding site (pCK4 MEC3), a COOH-terminal fragment of OB-cadherin not containing the juxtamembrane region (pCK4 OB-CAD CT), the entire cytoplasmic domain of Drosophila E-cadherin (pCK4 DEC), and smaller fragments of Drosophila E-cadherin as shown in B (pCK4 DEC 4, 14, and 15). (B) Schematic summary of the cadherin fragments and their interaction with δ-catenin. (C) Sequence alignment of the cytoplasmic tails of mouse E-cadherin, mouse OB-cadherin, and Drosophila DE-cadherin. Above the sequences are shown the smallest fragment of mouse E-cadherin which bound δ-catenin in the yeast two-hybrid assay, while below are diagrammed the amino acids missing from the OB-cadherin clone which does not bind to δ-catenin. The β-catenin/Armadillo binding site is also indicated.

Mentions: To determine whether the interaction between δ-catenin and the cytoplasmic domain of cadherin involves direct binding, their interaction was assayed in the yeast two-hybrid system. A construct was prepared containing the full Arm repeat region of δ-catenin, but lacking the NH2- and COOH-terminal domains. This construct specifically interacted with the full-length cytoplasmic tail of mouse E-cadherin (Fig. 7, A and B). Different classic cadherins in both mammals and flies carry two blocks of conserved sequence in their cytoplasmic tails (Fig. 7 C), a distal block which serves as the β-catenin/Armadillo binding site (Aberle et al., 1996; Pai et al., 1997) and a proximal region, which was recently discovered to bind p120ctn (Ozawa and Kemler, 1998; Yap et al., 1998). To begin to define the region required for δ-catenin binding, we tested two additional constructs, one containing the membrane-proximal region of mouse E-cadherin and the more distal region. δ-catenin specifically bound the membrane-proximal fragment, and failed to bind the more distal fragment. Also, δ-catenin did not bind to a truncated version of mouse OB-cadherin, which binds β-catenin (Tao et al., 1996), but lacks the 66 amino acids immediately following the transmembrane region that include the conserved proximal region (Fig. 7). We also tested whether δ-catenin could bind to Drosophila E-cadherin, to see whether the conserved sequences were sufficient for interaction. The full-length cytoplasmic tail of Drosophila E-cadherin binds δ-catenin, as do fragments containing the juxtamembrane region. The smallest interacting fragments of mouse E-cadherin and of Drosophila E-cadherin contain only the proximal 41 amino acids that include the highly conserved sequence YD(or E)D(or E)EGGGE (Fig. 7 C). This suggests that like p120ctn (Ozawa and Kemler, 1998; Yap et al., 1998), δ-catenin interacts with the conserved proximal region of the cadherin tail rather than the more distal site to which β-catenin and its fly homologue Armadillo bind (Aberle et al., 1996; Pai et al., 1997).


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)

δ-catenin interacts directly with the juxtamembrane region of cadherins in the yeast two-hybrid system. (A) Interaction of  δ-catenin with various cadherin fragments. The Arm repeat region of human δ-catenin in the pCK2 “bait” vector was tested against  pCK4 vector alone or with pCK4 fusions encoding the full-length murine E-cadherin cytoplasmic domain (pCK4 ME-CAD cyto), two  complementary fragments of murine E-cadherin carrying either the membrane-proximal region (pCK4 MEC2) or the distal region with  the β-catenin binding site (pCK4 MEC3), a COOH-terminal fragment of OB-cadherin not containing the juxtamembrane region  (pCK4 OB-CAD CT), the entire cytoplasmic domain of Drosophila E-cadherin (pCK4 DEC), and smaller fragments of Drosophila  E-cadherin as shown in B (pCK4 DEC 4, 14, and 15). (B) Schematic summary of the cadherin fragments and their interaction with  δ-catenin. (C) Sequence alignment of the cytoplasmic tails of mouse E-cadherin, mouse OB-cadherin, and Drosophila DE-cadherin.  Above the sequences are shown the smallest fragment of mouse E-cadherin which bound δ-catenin in the yeast two-hybrid assay, while  below are diagrammed the amino acids missing from the OB-cadherin clone which does not bind to δ-catenin. The β-catenin/Armadillo  binding site is also indicated.
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Related In: Results  -  Collection

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Figure 7: δ-catenin interacts directly with the juxtamembrane region of cadherins in the yeast two-hybrid system. (A) Interaction of δ-catenin with various cadherin fragments. The Arm repeat region of human δ-catenin in the pCK2 “bait” vector was tested against pCK4 vector alone or with pCK4 fusions encoding the full-length murine E-cadherin cytoplasmic domain (pCK4 ME-CAD cyto), two complementary fragments of murine E-cadherin carrying either the membrane-proximal region (pCK4 MEC2) or the distal region with the β-catenin binding site (pCK4 MEC3), a COOH-terminal fragment of OB-cadherin not containing the juxtamembrane region (pCK4 OB-CAD CT), the entire cytoplasmic domain of Drosophila E-cadherin (pCK4 DEC), and smaller fragments of Drosophila E-cadherin as shown in B (pCK4 DEC 4, 14, and 15). (B) Schematic summary of the cadherin fragments and their interaction with δ-catenin. (C) Sequence alignment of the cytoplasmic tails of mouse E-cadherin, mouse OB-cadherin, and Drosophila DE-cadherin. Above the sequences are shown the smallest fragment of mouse E-cadherin which bound δ-catenin in the yeast two-hybrid assay, while below are diagrammed the amino acids missing from the OB-cadherin clone which does not bind to δ-catenin. The β-catenin/Armadillo binding site is also indicated.
Mentions: To determine whether the interaction between δ-catenin and the cytoplasmic domain of cadherin involves direct binding, their interaction was assayed in the yeast two-hybrid system. A construct was prepared containing the full Arm repeat region of δ-catenin, but lacking the NH2- and COOH-terminal domains. This construct specifically interacted with the full-length cytoplasmic tail of mouse E-cadherin (Fig. 7, A and B). Different classic cadherins in both mammals and flies carry two blocks of conserved sequence in their cytoplasmic tails (Fig. 7 C), a distal block which serves as the β-catenin/Armadillo binding site (Aberle et al., 1996; Pai et al., 1997) and a proximal region, which was recently discovered to bind p120ctn (Ozawa and Kemler, 1998; Yap et al., 1998). To begin to define the region required for δ-catenin binding, we tested two additional constructs, one containing the membrane-proximal region of mouse E-cadherin and the more distal region. δ-catenin specifically bound the membrane-proximal fragment, and failed to bind the more distal fragment. Also, δ-catenin did not bind to a truncated version of mouse OB-cadherin, which binds β-catenin (Tao et al., 1996), but lacks the 66 amino acids immediately following the transmembrane region that include the conserved proximal region (Fig. 7). We also tested whether δ-catenin could bind to Drosophila E-cadherin, to see whether the conserved sequences were sufficient for interaction. The full-length cytoplasmic tail of Drosophila E-cadherin binds δ-catenin, as do fragments containing the juxtamembrane region. The smallest interacting fragments of mouse E-cadherin and of Drosophila E-cadherin contain only the proximal 41 amino acids that include the highly conserved sequence YD(or E)D(or E)EGGGE (Fig. 7 C). This suggests that like p120ctn (Ozawa and Kemler, 1998; Yap et al., 1998), δ-catenin interacts with the conserved proximal region of the cadherin tail rather than the more distal site to which β-catenin and its fly homologue Armadillo bind (Aberle et al., 1996; Pai et al., 1997).

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