TY - JOUR
T1 - Continual assembly of desmosomes within stable intercellular contacts of epithelial A-431 cells
AU - Gloushankova, Natalia A.
AU - Wakatsuki, Tetsuro
AU - Troyanovsky, Regina B.
AU - Elson, Elliot
AU - Troyanovsky, Sergey M.
N1 - Funding Information:
This work was supported by grant AR44016-04 from the National Institutes of Health
PY - 2003/12
Y1 - 2003/12
N2 - Subclones of human carcinoma-derived A-431 cell line stably producing fusion proteins consisting of the enhanced green fluorescent protein and either human desmoglein 2 (Dsg-GFP) or human plakoglobin (GFP-Pg) were used to examine the behavior of desmosomes in living cells. Immunofluorescence microscopy of the fixed cells showed that both fusion proteins, which were expressed in significantly lower levels relative to their endogenous counterparts, were efficiently recruited into desmosomes. Time-lapse confocal imaging of these cells reveals that such GFP-labeled desmosomes (GFP desmosomes) are stable structures which exhibit various dynamic and motile activities. The most notable are independent lateral mobility and fusion. Furthermore, the continual assembly of new nascent desmosomes is observed within stable contacts located at the middle of the epithelial sheet. A new GFP desmosome appears as a closely apposed group of fine patches which after a few minutes aggregate into a single structure. These three dynamic processes resulted in constant changes of desmosome distribution, numbers, and sizes. In addition, fluorescence recovery after photobleaching experiments showed that fine patches of desmosomal proteins may participate in desmosome maintenance. Such a diverse range of dynamic activities of desmosomes apparently produces flexible but tight cell-cell adhesion required for different morphogenetic events in epithelial structures.
AB - Subclones of human carcinoma-derived A-431 cell line stably producing fusion proteins consisting of the enhanced green fluorescent protein and either human desmoglein 2 (Dsg-GFP) or human plakoglobin (GFP-Pg) were used to examine the behavior of desmosomes in living cells. Immunofluorescence microscopy of the fixed cells showed that both fusion proteins, which were expressed in significantly lower levels relative to their endogenous counterparts, were efficiently recruited into desmosomes. Time-lapse confocal imaging of these cells reveals that such GFP-labeled desmosomes (GFP desmosomes) are stable structures which exhibit various dynamic and motile activities. The most notable are independent lateral mobility and fusion. Furthermore, the continual assembly of new nascent desmosomes is observed within stable contacts located at the middle of the epithelial sheet. A new GFP desmosome appears as a closely apposed group of fine patches which after a few minutes aggregate into a single structure. These three dynamic processes resulted in constant changes of desmosome distribution, numbers, and sizes. In addition, fluorescence recovery after photobleaching experiments showed that fine patches of desmosomal proteins may participate in desmosome maintenance. Such a diverse range of dynamic activities of desmosomes apparently produces flexible but tight cell-cell adhesion required for different morphogenetic events in epithelial structures.
KW - Cell culture
KW - Cell-cell adhesion
KW - Desmosomes
KW - Fluorescence recovery after photobleaching
KW - Green fluorescent protein
KW - Human
UR - http://www.scopus.com/inward/record.url?scp=0345117216&partnerID=8YFLogxK
U2 - 10.1007/s00441-003-0812-3
DO - 10.1007/s00441-003-0812-3
M3 - Article
C2 - 14564504
AN - SCOPUS:0345117216
SN - 0302-766X
VL - 314
SP - 399
EP - 410
JO - Cell and Tissue Research
JF - Cell and Tissue Research
IS - 3
ER -