cDNAs of cell adhesion molecules of different specificity induce changes in cell shape and border formation in cultured S180 cells.
The liver cell adhesion molecule (L-CAM) and N-cadherin or adherens junction-specific CAM (A-CAM) are structurally related cell surface glycoproteins that mediate calcium-dependent adhesion in different tissues. We have isolated and characterized a full-length cDNA clone for chicken N-cadherin and used this clone to transfect S180 mouse sarcoma cells that do not normally express N-cadherin. The transfected cells (S180cadN cells) expressed N-cadherin on their surfaces and resembled S180 cells transfected with L-CAM (S180L cells) in that at confluence they formed an epithelioid sheet and displayed a large increase in the number of adherens and gap junctions. In addition, N-cadherin in S180cadN cells, like L-CAM in S180L cells, accumulated at cellular boundaries where it was colocalized with cortical actin. In S180L cells and S180cadN cells, L-CAM and N-cadherin were seen at sites of adherens junctions but were not restricted to these areas. Adhesion mediated by either CAM was inhibited by treatment with cytochalasin D that disrupted the actin network of the transfected cells. Despite their known structural similarities, there was no evidence of interaction between L-CAM and N-cadherin. Doubly transfected cells (S180L/cadN) also formed epithelioid sheets. In these cells, both N-cadherin and L-CAM colocalized at areas of cell contact and the presence of antibodies to both CAMs was required to disrupt the sheets of cells. Studies using divalent antibodies to localize each CAM at the cell surface or to perturb their distributions indicated that in the same cell there were no interactions between L-CAM and N-cadherin molecules. These data suggest that the Ca(++)-dependent CAMs are likely to play a critical role in the maintenance of epithelial structures and support a model for the segregation of CAM mediated binding. They also provide further support for the so-called precedence hypothesis that proposes that expression and homophilic binding of CAMs are necessary for formation of junctional structures in epithelia.
Matsuzaki
F
F
Rockefeller University, New York 10021.
Mège
R M
RM
Jaffe
S H
SH
Friedlander
D R
DR
Gallin
W J
WJ
Goldberg
J I
JI
Cunningham
B A
BA
Edelman
G M
GM
D000818:Animals; D015820:Cadherins / Q000235:genetics* / Q000502:physiology; D002118:Calcium / Q000494:pharmacology; D002448:Cell Adhesion / Q000187:drug effects; D015815:Cell Adhesion Molecules / Q000235:genetics* / Q000502:physiology; D002642:Chick Embryo; D015638:Cytochalasin D / Q000494:pharmacology; D004247:DNA / Q000235:genetics; D005455:Fluorescent Antibody Technique; D015723:Gene Library; D007365:Intercellular Junctions / Q000648:ultrastructure; D051379:Mice; D008854:Microscopy, Electron; D015183:Restriction Mapping; D012510:Sarcoma 180 / Q000473:pathology; D014162:Transfection; D014407:Tumor Cells, Cultured / Q000166:cytology* / Q000648:ultrastructure
D015820:Cadherins; D015815:Cell Adhesion Molecules; D015638:Cytochalasin D; D004247:DNA; D002118:Calcium