Identification of amino acid sequence motifs in desmocollin, a desmosomal glycoprotein, that are required for plakoglobin binding and plaque formation.
By transfecting epithelial cells with gene constructs encoding chimeric proteins of the transmembrane part of the gap junction protein connexin 32 in combination with various segments of the cytoplasmic part of the desmosomal cadherin desmocollin 1a, we have determined that a relatively short sequence element is necessary for the formation of desmosome-like plaques and for the specific anchorage of bundles of intermediate-sized filaments (IFs). Deletion of as little as the carboxyl-terminal 37 aa resulted in a lack of IF anchorage and binding of the plaque protein plakoglobin, as shown by immunolocalization and immunoprecipitation experiments. In addition, we show that the sequence requirements for the recruitment of desmoplakin, another desmosomal plaque protein, differ and that a short (10 aa) segment of the desmocollin 1a tail, located close to the plasma membrane, is also required for the binding of plakoglobin, as well as of desmoplakin, and also for IF anchorage. The importance of the carboxyl-terminal domain, homologous in diverse types of cadherins, is emphasized, as it must harbor, in a mutually exclusive pattern, the information for assembly of the IF-anchoring desmosomal plaque in desmocollins and for formation of the alpha-/beta-catenin- and vinculin-containing, actin filament-anchoring plaque in E- and N-cadherin.
Division of Cell Biology, German Cancer Research Center, Heidelberg.
S M
SM
Troyanovsky
R B
RB
Troyanovsky
L G
LG
Eshkind
R E
RE
Leube
W W
WW
Franke
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002417:Cattle; D002448:Cell Adhesion; D017630:Connexins / Q000737:chemistry; D003598:Cytoskeletal Proteins / Q000737:chemistry* / Q000378:metabolism*; D051187:Desmocollins; D051180:Desmoplakins; D003896:Desmosomes / Q000648:ultrastructure*; D007381:Intermediate Filament Proteins / Q000378:metabolism; D007382:Intermediate Filaments / Q000648:ultrastructure; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D011485:Protein Binding; D051381:Rats; D011993:Recombinant Fusion Proteins; D013329:Structure-Activity Relationship; D051185:gamma Catenin; D000097002:Gap Junction beta-1 Protein
D017630:Connexins; D003598:Cytoskeletal Proteins; D051187:Desmocollins; D051180:Desmoplakins; D007381:Intermediate Filament Proteins; D009838:Oligodeoxyribonucleotides; D011993:Recombinant Fusion Proteins; D051185:gamma Catenin; D000097002:Gap Junction beta-1 Protein