Defining the interactions between intermediate filaments and desmosomes.
Desmoplakin (DP), plakoglobin (PG), and plakophilin 1 (PP1) are desmosomal components lacking a transmembrane domain, thus making them candidate linker proteins for connecting intermediate filaments and desmosomes. Using deletion and site-directed mutagenesis, we show that remarkably, removal of approximately 1% of DP's sequence obliterates its ability to associate with desmosomes. Conversely, when linked to a foreign protein, as few as 86 NH2-terminal DP residues are sufficient to target to desmosomes efficiently. In in vitro overlay assays, the DP head specifically associates with itself and with desmocollin 1a (Dsc1a). In similar overlay assays, PP1 binds to DP and Dsc1a, and to a lesser extent, desmoglein 1 (Dsg1), while PG binds to Dsg1 and more weakly to Dsc1a and DP. Interestingly, like DP, PG and PP1 associate with epidermal keratins, although PG is considerably weaker in its ability to do so. As judged by overlay assays, the amino terminal head domain of type II keratins appears to have a special importance in establishing these connections. Taken together, our findings provide new insights into the complexities of the links between desmosomes and intermediate filaments (IFs). Our results suggest a model whereby at desmosome sites within dividing epidermal cells, DP and PG anchor to desmosomal cadherins and to each other, forming an ordered array of nontransmembrane proteins that then bind to keratin IFs. As epidermal cells differentiate, PP1 is added as a molecular reinforcement to the plaque, enhancing anchorage to IFs and accounting at least partially for the increase in numbers and stability of desmosomes in suprabasal cells.
Smith
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Howard Hughes Medical Institute, The University of Chicago, Chicago, Illinois 60637, USA.
Fuchs
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D000595:Amino Acid Sequence; D001665:Binding Sites; D002478:Cells, Cultured; D003598:Cytoskeletal Proteins / Q000235:genetics / Q000378:metabolism*; D051187:Desmocollins; D051183:Desmoglein 1; D051182:Desmogleins; D051180:Desmoplakins; D003896:Desmosomes / Q000378:metabolism*; D004817:Epidermis; D006801:Humans; D007382:Intermediate Filaments / Q000378:metabolism*; D015603:Keratinocytes; D007633:Keratins / Q000378:metabolism; D008969:Molecular Sequence Data; D016296:Mutagenesis; D051191:Plakophilins; D011506:Proteins / Q000378:metabolism; D051185:gamma Catenin
D003598:Cytoskeletal Proteins; C495297:DSG1 protein, human; C495294:DSP protein, human; D051187:Desmocollins; D051183:Desmoglein 1; D051182:Desmogleins; D051180:Desmoplakins; C495335:PKP1 protein, human; D051191:Plakophilins; D011506:Proteins; D051185:gamma Catenin; D007633:Keratins