Desmocollin-2 alone forms functional desmosomal plaques, with the plaque formation requiring the juxtamembrane region and plakophilins
العنوان: | Desmocollin-2 alone forms functional desmosomal plaques, with the plaque formation requiring the juxtamembrane region and plakophilins |
---|---|
المؤلفون: | Tetsushi Sakuma, Megumi Tsuda, Miwako Fujiwara, Shuichi Obata, Shintaro T. Suzuki, Azusa Nagatomo, Takashi Yamamoto |
المصدر: | Journal of Biochemistry. 158:339-353 |
بيانات النشر: | Oxford University Press (OUP), 2015. |
سنة النشر: | 2015 |
مصطلحات موضوعية: | Immunoprecipitation, Recombinant Fusion Proteins, Desmoglein-2, Biology, Biochemistry, Microscopy, Electron, Transmission, Antigens, CD, Desmosome, Mutant protein, Cell Line, Tumor, Cell Adhesion, medicine, Humans, Protein Interaction Domains and Motifs, Cell adhesion, Molecular Biology, Gene knockout, Desmocollins, Desmoglein 2, Epithelial Cells, Desmosomes, General Medicine, Cadherins, Molecular biology, Peptide Fragments, Recombinant Proteins, medicine.anatomical_structure, Microscopy, Fluorescence, Mutant Proteins, Desmocollin, CRISPR-Cas Systems, Plakophilins, Gene Deletion |
الوصف: | The role of the juxtamembrane region of the desmocollin-2 cytoplasmic domain in desmosome formation was investigated by using gene knockout and reconstitution experiments. When a deletion construct of the desmocollin-2 juxtamembrane region was expressed in HaCaT cells, the mutant protein became localized linearly at the cell-cell boundary, suggesting the involvement of this region in desmosomal plaque formation. Then, desmocollin-2 and desmoglein-2 genes of epithelial DLD-1 cells were ablated by using the CRISPR/Cas9 system. The resultant knockout cells did not form desmosomes, but re-expression of desmocollin-2 in the cells formed desmosomal plaques in the absence of desmoglein-2 and the transfectants showed significant cell adhesion activity. Intriguingly, expression of desmocollin-2 lacking its juxtamembrane region did not form the plaques. The results of an immunoprecipitation and GST-fusion protein pull-down assay suggested the binding of plakophilin-2 and -3 to the region. Ablation of plakophilin-2 and -3 genes resulted in disruption of the plaque-like accumulation and linear localization of desmocollin-2 at intercellular contact sites. These results suggest that the juxtamembrane region of desmocollin-2 and plakophilins are involved in the desmosomal plaque formation, possibly through the interaction between this region and plakophilins. |
تدمد: | 1756-2651 0021-924X |
URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::63056852b86e64e61219a5e83eb0a5f0 https://doi.org/10.1093/jb/mvv048 |
رقم الأكسشن: | edsair.doi.dedup.....63056852b86e64e61219a5e83eb0a5f0 |
قاعدة البيانات: | OpenAIRE |
تدمد: | 17562651 0021924X |
---|