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Medical Research Council Protein Phosphorylation Unit, Department of Biochemistry, University of Dundee, Dundee, United Kingdom
LPS stimulation of RAW264 macrophages triggered the activation of
mitogen- and stress-activated protein kinases-1 and -2 (MSK1, MSK2) and
their putative substrates, the transcription factors cyclic AMP
response element-binding protein (CREB) and activating transcription
factor-1 (ATF1). The activation of MSK1/MSK2 was prevented by
preincubating the cells with a combination of two drugs that suppress
activation of the classical mitogen-activated protein kinase cascade
and stress-activated protein kinase/p38, respectively, but inhibition
was only partial in the presence of either inhibitor. The
LPS-stimulated activation of CREB and ATF1, the transcription of the
cyclooxygenase-2 (COX-2) and IL-1ß genes (the promoters of which
contain a cyclic AMP response element), and the induction of the COX-2
protein were prevented by the same drug combination, as well as by Ro
318220 or H89, potent inhibitors of MSK1/MSK2. Two other transcription
factors, C/EBPß and NF-
B, have been implicated in the
transcription of the COX-2 gene. However, PD 98059 and/or SB 203580 did
not prevent the LPS-induced increase in the level of the transcription
factor C/EBPß, and none of the four inhibitors used in this study
prevented the activation of NF-B. Our results demonstrate that two
different mitogen-activated protein kinase cascades are rate limiting
for the LPS-induced activation of CREB/ATF1 and the transcription of
the COX-2 and IL-1ß genes. They also suggest that MSK1 and MSK2 may
play a role in these processes and hence are potential targets for the
development of novel antiinflammatory drugs.
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