This application is for continued support of our studies of the molecular regulation of the acute phase pentraxin C reactive protein (CRP), its homologue serum amyloid P component (SAP) and the related acute phase protein serum amyloid A (SAA). The availability of cDNA and genomic clones generated during the initial grant period provides the reagents to pursue the proposed studies of mechanisms of pentraxin and other acute phase gene regulation and its relationship to the pathophysiology of amyloidosis. Specifically, we plan to: 1) complete characterization of the human pentraxin genes and compare them to the corresponding hamster genes. 2) Determine the mechanisms involved in pentraxin and SAA gene expression by defining the mediators and the structure and organization of promoter and enhancer sequence flanking these genes. 3) Examine the basis for hepatic and extrahepatic expression of SAA. To accomplish these goals we will use phage and cosmid cloning methods, transient and stable transfection systems, in vitro transcription and translation analysis and in situ hybridization techniques. The availability of an excellent model of amyloidosis in the Syrian hamster and cDNA clones for each of the relevant proteins permit comparisons of these genes to the corresponding human genes and gene products. A basis for understanding of the amyloidogenic process resulting from acute phase stimuli should result. The proposed studies will have general implications for understanding mechanisms of eukaryotic gene control as well as questions specifically related to cytokine mediated control of inflammation.
