Identification of the plakoglobin-binding domain in desmoglein and its role in plaque assembly and intermediate filament anchorage.
The carboxyterminal cytoplasmic portions (tails) of desmosomal cadherins of both the desmoglein (Dsg) and desmocollin type are integral components of the desmosomal plaque and are involved in desmosome assembly and the anchorage of intermediate-sized filaments. When additional Dsg tails were introduced by cDNA transfection into cultured human epithelial cells, in the form of chimeras with the aminoterminal membrane insertion domain of rat connexin32 (Co32), the resulting stably transfected cells showed a dominant-negative defect specific for desmosomal junctions: despite the continual presence of all desmosomal proteins, the endogenous desmosomes disappeared and the formation of Co32-Dsg chimeric gap junctions was inhibited. Using cell transfection in combination with immunoprecipitation techniques, we have examined a series of deletion mutants of the Dsg1 tail in Co32-Dsg chimeras. We show that upon removal of the last 262 amino acids the truncated Dsg tail still effects the binding of plakoglobin but not of detectable amounts of any catenin and induces the dominant-negative phenotype. However, further truncation or excision of the next 41 amino acids, which correspond to the highly conserved carboxyterminus of the C-domain in other cadherins, abolishes plakoglobin binding and allows desmosomes to reform. Therefore, we conclude that this short segment provides a plakoglobin-binding site and is important for plaque assembly and the specific anchorage of either actin filaments in adherens junctions or IFs in desmosomes.
Troyanovsky
S M
SM
Division of Cell Biology, Germany Cancer Research Center, Heidelberg.
Troyanovsky
R B
RB
Eshkind
L G
LG
Krutovskikh
V A
VA
Leube
R E
RE
Franke
W W
WW
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002448:Cell Adhesion / Q000502:physiology; D017630:Connexins / Q000235:genetics / Q000378:metabolism; D003598:Cytoskeletal Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D051187:Desmocollins; D051183:Desmoglein 1; D051182:Desmogleins; D051180:Desmoplakins; D003896:Desmosomes / Q000378:metabolism*; D004847:Epithelial Cells; D006801:Humans; D007382:Intermediate Filaments / Q000378:metabolism*; D008969:Molecular Sequence Data; D011233:Precipitin Tests; D017434:Protein Structure, Tertiary; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D016415:Sequence Alignment; D017384:Sequence Deletion / Q000502:physiology; D014162:Transfection; D014407:Tumor Cells, Cultured; D051185:gamma Catenin; D000097002:Gap Junction beta-1 Protein
D017630:Connexins; D003598:Cytoskeletal Proteins; D051187:Desmocollins; D051183:Desmoglein 1; D051182:Desmogleins; D051180:Desmoplakins; D011993:Recombinant Fusion Proteins; D051185:gamma Catenin; D000097002:Gap Junction beta-1 Protein; C495297:DSG1 protein, human