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J Biol Chem, Vol. 274, Issue 15, 10140-10144, April 9, 1999
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From the Department of Adult Oncology, Dana-Farber Cancer
Institute, Harvard Medical School, Boston, Massachusetts 02115, The stress-activated p38 mitogen-activated
protein kinase (p38 MAPK), a member of the subgroup of mammalian
kinases, appears to play an important role in regulating inflammatory
responses, including cytokine secretion and apoptosis. The upstream
mediators that link extracellular signals with the p38 MAPK signaling
pathway are currently unknown. Here we demonstrate that pp125 focal
adhesion kinase-related tyrosine kinase RAFTK (also known as PYK2,
CADTK) is activated specifically by methylmethane sulfonate (MMS) and hyperosmolarity but not by ultraviolet radiation, ionizing radiation, or cis-platinum. Overexpression of RAFTK leads to the
activation of p38 MAPK. Furthermore, overexpression of a
dominant-negative mutant of RAFTK (RAFTK K-M) inhibits MMS-induced p38
MAPK activation. MKK3 and MKK6 are known potential constituents of p38
MAPK signaling pathway, whereas SEK1 and MEK1 are upstream activators
of SAPK/JNK and ERK pathways, respectively. We observe that the
dominant-negative mutant of MKK3 but not of MKK6, SEK1, or MEK1
inhibits RAFTK-induced p38 MAPK activity. Furthermore, the results
demonstrate that treatment of cells with
1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid, tetra(acetoxymethyl)-ester, a membrane-permeable calcium chelator, inhibits MMS-induced activation of RAFTK and p38 MAPK. Taken
together, these findings indicate that RAFTK represents a
stress-sensitive mediator of the p38 MAPK signaling pathway in response
to certain cytotoxic agents.
Division of Experimental Medicine and Hematology,
Deaconess Medical Center, Harvard Institutes of Medicine, Boston,
Massachusetts 02115, § Department of Molecular Biology,
Diabetes Research Laboratory, Massachusetts General Hospital, Boston,
Massachusetts 02114, and ¶ Department of Biochemistry and
Molecular Biology, Howard Hughes Medical Institute, Program in
Molecular Medicine, University of Massachusetts Medical School,
Worcester, Massachusetts 01605
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