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J Biol Chem, Vol. 274, Issue 29, 20185-20190, July 16, 1999
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From the Hypertonicity induces a group of genes that are
responsible for the intracellular accumulation of protective organic
osmolytes such as sorbitol and betaine. Two representative genes are
the aldose reductase enzyme (AR, EC 1.1.1.21), which is responsible for
the conversion of glucose to sorbitol, and the betaine transporter (BGT1), which mediates Na+-coupled betaine uptake in
response to osmotic stress. We recently reported that the induction of
BGT1 mRNA in the renal epithelial Madin-Darby canine kidney cell
line is inhibited by SB203580, a specific p38 kinase inhibitor. In
these studies we report that the hypertonic induction of aldose
reductase mRNA in HepG2 cells as well as the osmotic response
element (ORE)-driven reporter gene expression in transfected HepG2
cells are both inhibited by SB203580, suggesting that p38 kinase
mediates the activation and/or binding of the transcription factor(s)
to the ORE. Electrophoretic gel mobility shift assays with cell
extracts prepared from SB203580-treated, hypertonically stressed HepG2
cells further show that the binding of trans-acting factors to the ORE
is prevented and is thus also dependent on the activity of p38 kinase.
Similarly, treatment of hypertonically stressed cells with PD098059, a
mitogen-activated extracellular regulated kinase kinase (MEK1)
inhibitor, results in inhibition of the hypertonic induction of aldose
reductase mRNA, ORE-driven reporter gene expression, and the
binding of trans-acting factors to the ORE. ORE-driven reporter gene
expression was not affected by p38 kinase inhibition or MEK1 inhibition
in cells incubated in iso-osmotic media. These data indicate that p38
kinase and MEK1 are involved in the regulation of the hyperosmotic stress response.
Harry B. and Aileen Gordon Diabetes Research
Laboratory,
Copyright © 1999 by The American Society for Biochemistry and Molecular Biology, Inc.
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