The results from numerous studies indicate that interleukin 1 (IL 1), a low molecular weight protein produced by stimulated macrophages and perhaps several other cell types, can markedly affect the maturation, functional activation, and proliferation of a broad range of cell types that share a common involvement immune or inflammatory responses. In view of the proposed role of IL 1 in immune and inflammation, it is important to understand the mechanism of action of the mediator at the biochemical and molecular levels. Thus the overall goal of our proposed studies is to develop an understanding the events that follow the binding of IL 1 to its receptor that ultimately trigger the activation of specific genes, for example, the interleukin 2 receptor in lymphocytes and kappa immunoglobulin light chain in pre-B cells. Our recent studies indicate that cAMP may be the intracellular second messenger for IL 1 action. We plan, therefore, to focus our attention on the molecular event that are associated with the activation of adenyl cyclase by IL 1 as well as the mechanism by which IL 1 triggers gene activation. In the former case, we will determine if IL 1 activates adenyl cyclase via a GTP-binding protein intermediate. In the gene activation studies, we will determine if IL 1 induces the activation specific DNA-binding proteins with specificity for regulatory sequences in the gene for the IL 2 receptor and kappa immunoglobulin light chain. We will characterize IL 1-responsive DNA sequences that are associated with activation of these gene The studies on CAMP and gene activation will be linked by studies on the possible role of cAMP-dependent protein kinases in the activation of specific DNA-binding proteins. In the work outlined in this proposal , we will depend on the use of standard methods in recombinant DNA technology (for example, construction of plasmids, transient expression assays, and DNA mobility shift and methylation interference assays) as well as well-established biochemical procedures for the study of cAMP metabolism and action.
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