Focal adhesion kinase regulates intestinal epithelial barrier function via redistribution of tight junction.
Disruption of epithelial barrier function was identified as one of the pathologic mechanisms in inflammatory bowel diseases (IBD). Epithelial barrier consists of various intercellular junctions, in which the tight junction (TJ) is an important component. However, the regulatory mechanism of tight junction is still not clear. Here we examined the role of focal adhesion kinase (FAK) in the epithelial barrier function on Caco-2 monolayers using a specific FAK inhibitor, PF-573, 228 (PF-228). We found that the decrease of transepithelial resistance and the increase of paracellular permeability were accompanied with the inhibition of autophosphorylation of FAK by PF-228 treatment. In addition, PF-228 inhibited the FAK phosphorylation at Y576/577 on activation loop by Src, suggesting Src-dependent regulation of FAK in Caco-2 monolayers. In an ethanol-induced barrier injury model, PF-228 treatment also inhibited the recovery of transepithelial resistance as well as these phosphorylations of FAK. In a sucrose gradient ultracentrifugation, FAK co-localized with claudin-1, an element of the TJ complex, and they co-migrate after ethanol-induced barrier injury. Immunofluorescence imaging analysis revealed that PF-228 inhibited the FAK redistribution to the cell border and reassembly of TJ proteins in the recovery after ethanol-induced barrier injury. Finally, knockdown of FAK by siRNA resulted in the decrease of transepithelial resistance. These findings reveal that activation of FAK is necessary for maintaining and repairing epithelial barrier in Caco-2 cell monolayer via regulating TJ redistribution.
Department of Biochemistry, Asahikawa Medical University, Asahikawa, Japan.
Yanju
Y
Ma
Shingo
S
Semba
Rafiqul Islam
RI
Khan
Hiroki
H
Bochimoto
Tsuyoshi
T
Watanabe
Mikihiro
M
Fujiya
Yutaka
Y
Kohgo
Yunpeng
Y
Liu
Takanobu
T
Taniguchi
D020816:Amino Acid Motifs; D018938:Caco-2 Cells; D004847:Epithelial Cells / Q000201:enzymology*; D051416:Focal Adhesion Protein-Tyrosine Kinases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007422:Intestines / Q000166:cytology / Q000201:enzymology*; D010766:Phosphorylation; D019108:Tight Junctions / Q000201:enzymology* / Q000235:genetics
D051416:Focal Adhesion Protein-Tyrosine Kinases