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J. Biol. Chem., Vol. 275, Issue 22, 16933-16940, June 2, 2000
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From the Departments of A member of the family of p21-activated protein
kinases,
p21-activated Protein Kinase
-PAK Is Translocated and
Activated in Response to Hyperosmolarity
IMPLICATION OF Cdc42 AND PHOSPHOINOSITIDE 3-KINASE IN A TWO-STEP
MECHANISM FOR 
-PAK ACTIVATION*
,,¶
Biochemistry and
§ Biomedical Sciences, University of California,
Riverside, California 92521
-PAK, has cytostatic properties and is activated during
apoptosis and in response to DNA damage. To determine whether -PAK
is activated by other types of cell stress and to assess its mechanism
of activation, the response of -PAK to hyperosmotic stress was
examined. In 3T3-L1 mouse fibroblasts, there are two pools of -PAK:
the majority of the protein kinase is soluble and has low specific
activity, whereas -PAK associated with the particulate fraction has
significantly higher specific activity. Hyperosmolarity promotes
translocation of -PAK from the soluble to the particulate fraction;
this parallels activation of the protein kinase. Activation but not
translocation of -PAK is wortmannin-sensitive, suggesting the
involvement of a phosphoinositide 3-kinase-related activity. -PAK
translocation in response to hyperosmolarity parallels Cdc42
translocation to the particulate fraction in vivo and can
be induced in vitro by guanosine
5'-3-O-(thio)triphosphate. Cotransfection of -PAK with constitutively active Cdc42 induces -PAK activation and
translocation, whereas inactive Cdc42 inhibits both processes in
response to hyperosmotic stress, suggesting that Cdc42 has a role in
the translocation and activation of -PAK. 
-PAK is not activated
in response to hyperosmolarity in 3T3-L1 cells. A two-step model of
-PAK activation is presented.
*
This research was supported by United States Public Health
Service Grant GM26738 (to J. A. T.).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.
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