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Is Attenuated in Murine Aortic Endothelial Cells Derived from Double-Stranded RNA-Activated Kinase (PKR)-Null Mice1
Departments of
*
Cancer Biology,
Urology, and
Colorectal Surgery, Lerner Research Institute, Cleveland Clinic Foundation, Cleveland, OH 44195
The adherence of leukocytes on the endothelium is mediated in part
by the transient expression of the E-selectin adhesion molecule.
Because we have previously shown that the dsRNA-activated kinase PKR
mediates dsRNA induction of NF-
B, we used murine aortic endothelial
(MuAE) cells isolated from wild-type and PKR-null mice to investigate
the role of PKR in the induction of E-selectin expression by dsRNA
(pIC) and TNF-
. E-selectin mRNA and protein expression was inducible
by both pIC and TNF- in wild-type MuAE cells, whereas induction of
E-selectin expression by these agents was defective in PKR-null MuAE
cells. Induction of E-selectin promoter activity and NF-B DNA
binding activity were substantially reduced in pIC- or TNF--treated
PKR-null cells, indicating a role for PKR in both pIC and TNF-
induction of E-selectin via an NF-B-dependent pathway. In PKR-null
cells, pIC-mediated degradation of IBß is deficient. Activation of
this pathway requires the PKR-dependent degradation of the IBß
protein. Moreover, both phosphorylated and unphosphorylated activating
transcription factor 2 DNA-binding activities were reduced in PKR-null
aortic endothelial cells. These results indicate that the PKR is
required for full activation of E-selectin expression by pIC and
TNF- in primary mouse aortic endothelial cells identifying
activating transcription factor 2 as a new target for PKR-dependent
regulation and suggest a role for PKR in leukocyte
adhesion.
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