ISSN:
0886-1544
Keywords:
adhering junctions
;
desmosome
;
assembly
;
phosphorylation
;
protein interaction
;
Life and Medical Sciences
;
Cell & Developmental Biology
Source:
Wiley InterScience Backfile Collection 1832-2000
Topics:
Biology
,
Medicine
Notes:
We have analyzed the kinetics of synthesis, phosphorylation, and stability of the soluble and insoluble plakoglobin (PG) and their interactions with Dsg1 and E-cadherin in Madin-Darby canine kidney (MDCK) epithelial cells in the absence of cell adhesion and after the induction of cell-cell contact. Using a combination of biochemical and morphological approaches, we show that newly synthesized PG enters a soluble:insoluble pool of proteins in a 60:40 ratio regardles of cell-cell contact. Following synthesis, PG is increasingly found in the insoluble pool. Although cell-cell contact does not effect either the size of each pool or the rate or efficiency of the transfer from the soluble into the insoluble pool, it results in a significant increase in the metabolic stability of the newly synthesized insoluble PG. The soluble PG initially forms separate complexes with E-cadherin and Dsg1. PG-Dsg1 complexes become insoluble and localize to the desmosome. PG-E-cadherin complexes remain soluble and are distributed intracellularly. The insoluble PG and E-cadherin detected at the cell periphery remain distinctly separate, as demonstrated previously [Hinck et al., 1994: J. Cell Biol. 125:1327-1340; Nathke et al., 1994: J. Cell Biol. 125:1341-1352]. In addition, we detected a separate pool of PG which is not associated with either Dsg1 or E-cadherin and after the induction of cell-cell contact becomes primarily insoluble and is distributed along the lateral membrane. Phoshorylation analysis showed that there is a significantly greater amount of phosphorylated PG in the soluble pool than in the insoluble pool. In addition the soluble pool is both serine and theronine phosphorylated, whereas the insoluble PG is primarily phosphorylated on serine residues. © 1995 Wiley-Liss, Inc.
Additional Material:
5 Ill.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1002/cm.970320403
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