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J. Biol. Chem., Vol. 276, Issue 39, 36183-36193, September 28, 2001
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From the Decreased phosphorylation of focal adhesion
kinase (FAK) is associated with loss of focal adhesions and actin
stress fibers and precedes the onset of apoptosis in renal epithelial
cells caused by nephrotoxicants (Van de Water, B., Nagelkerke, J. F., and Stevens, J. L. (1999) J. Biol. Chem. 274, 13328-13337). The role of FAK in the control of apoptosis caused by
nephrotoxicants was further investigated in LLC-PK1 cells that were
stably transfected with either green fluorescent protein (GFP)-FAK or
dominant negative acting deletion mutants of FAK, GFP-FAT, and
GFP-FRNK. GFP-FAT and GFP-FRNK delayed the formation of focal adhesions
and prevented the localization of endogenous (phosphorylated) FAK at
these sites. GFP-FAT and GFP-FRNK overexpression potentiated the onset
of apoptosis caused by the nephrotoxicant dichlorovinyl-cysteine. This
was associated with an increased activation of caspase-3. GFP-FAT also
potentiated apoptosis caused by doxorubicin but not cisplatin. The
potentiation of apoptosis by GFP-FAT was related to an almost complete
dephosphorylation of FAK; this did not occur in cells overexpressing
only GFP. This dephosphorylation was associated with a pronounced loss
of focal adhesion organization in GFP-FAT cells, in association with
loss of tyrosine phosphorylation of paxillin. In conclusion, the data
indicate an important role of cell-matrix signaling in the control of
chemically induced apoptosis; loss of FAK activity caused by toxic
chemicals results in perturbations of focal adhesion organization with
a subsequent inactivation of associated (signaling) molecules and loss
of survival signaling.
Suppression of Chemically Induced Apoptosis but Not Necrosis of
Renal Proximal Tubular Epithelial (LLC-PK1) Cells by Focal Adhesion
Kinase (FAK)
ROLE OF FAK IN MAINTAINING FOCAL ADHESION ORGANIZATION AFTER
ACUTE RENAL CELL INJURY*
§,,, and
Division of Toxicology, Leiden Amsterdam
Center for Drug Research, Leiden University,
2300 RA Leiden, The Netherlands
*
This work was supported by Grants 902-21-208 and 902-21-217 from the Dutch Organization for Scientific Research and a fellowship from the Royal Netherlands Academy for Arts and Sciences (to
B. v. d. W.).The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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