A genome-wide screen identifies conserved protein hubs required for cadherin-mediated cell-cell adhesion.
Cadherins and associated catenins provide an important structural interface between neighboring cells, the actin cytoskeleton, and intracellular signaling pathways in a variety of cell types throughout the Metazoa. However, the full inventory of the proteins and pathways required for cadherin-mediated adhesion has not been established. To this end, we completed a genome-wide (~14,000 genes) ribonucleic acid interference (RNAi) screen that targeted Ca(2+)-dependent adhesion in DE-cadherin-expressing Drosophila melanogaster S2 cells in suspension culture. This novel screen eliminated Ca(2+)-independent cell-cell adhesion, integrin-based adhesion, cell spreading, and cell migration. We identified 17 interconnected regulatory hubs, based on protein functions and protein-protein interactions that regulate the levels of the core cadherin-catenin complex and coordinate cadherin-mediated cell-cell adhesion. Representative proteins from these hubs were analyzed further in Drosophila oogenesis, using targeted germline RNAi, and adhesion was analyzed in Madin-Darby canine kidney mammalian epithelial cell-cell adhesion. These experiments reveal roles for a diversity of cellular pathways that are required for cadherin function in Metazoa, including cytoskeleton organization, cell-substrate interactions, and nuclear and cytoplasmic signaling.
Department of Biology and 2 Department of Molecular and Cellular Physiology, Stanford University, Stanford, CA 94305.
Christopher P
CP
Toret
Michael V
MV
D'Ambrosio
Ronald D
RD
Vale
Michael A
MA
Simon
W James
WJ
Nelson
D000818:Animals; D015820:Cadherins / Q000378:metabolism / Q000502:physiology*; D002448:Cell Adhesion / Q000235:genetics* / Q000502:physiology; D002478:Cells, Cultured; D004285:Dogs; D029721:Drosophila Proteins / Q000235:genetics* / Q000502:physiology; D004331:Drosophila melanogaster / Q000166:cytology / Q000378:metabolism*; D055785:Gene Knockdown Techniques; D049750:Genome, Insect; D061985:Madin Darby Canine Kidney Cells; D034622:RNA Interference
D015820:Cadherins; D029721:Drosophila Proteins