A multi-user group proposes to purchase a phosphorimager for the rapid, sensitive, and accurate detection and quantitation of radioactive and chemiluminescent species in analyses of the molecular genetics of signal transduction, DNA replication, and gene expression. Specific projects include analysis of the mechanism by which DNA replication is initiated in eukaryotic cells and how this process is regulated; the mechanism of activation of Ras proteins by receptor tyrosine kinases and the identification of downstream targets of activated Ras proteins; the dissection of protein-protein interactions that regulate transcription of genes in eukaryotic cells by RNA polymerase II; testing models for the structure of large RNA molecules and determination of the ways in which RNA structure can influence gene expression, and exploration of signal transduction pathways involved in cell growth and development. Specific applications that will utilize the phosphorimager include in vivo radiolabelling of proteins; determination of protein half-life; analysis of protein posttranslational modificatimns, subcellular localization and oligomeric states; analysis of protein-protein, protein-nucleic acid, and intranucleic acid interactions; analysis of transcriptional control elements; analysis of in vivo and in vitro DNA replication products. The results of these studies will significantly extend our understanding of basic cellular processes.
