The proposal before you requests partial funding for a PhosphorImager to be used by members of the Departments of Pharmacology and Molecular Cancer Biology of the Duke University Medical Center. The proposal describes two major areas of research that join the different investigators (both major users and minor users), in common pursuit. First, is the understanding of signal transduction networks in eukaryotic organisms, particularly in areas related to the cell cycle and cellular differentiation. Second, is the understanding of the structure, function and regulation of eukaryotic genes. The elucidation of the networks of intracellular signalling in eukaryotes will be accomplished by work on a variety of organisms. The work of Drs. Garrett and Lew addresses issues of cell cycle regulation and differentiation in the baker's yeast Saccharomyces cerevisiae. Dr. Means uses Aspergillus nidulans to address the importance of calcium binding proteins in signalling. Xenopous laevis oocytes and mammalian cell lines are used by Drs. Bell, Pendergast, Shenolikar, Swenson and Wang to elucidate signalling pathways with an emphasis on the importance of phosphorylation and dephosphorylation. Dr. Casey also uses mammalian cell lines to unravel the signalling mediated by small G proteins,. The precise regulation of intracellular signalling is modulated by subtle changes in multiple pathways. Reliable quantification of these changes has been difficult, if not impossible, to document using autoradiographic techniques. PhosphorImager technology is the standard for this quantification and the studies described in the proposal require ready acces to this technology. The study of gene structure and expression has resulted in some of the most dramatic revolutions in modern biology. Work to understand the transcription regulation of gene expression following cellular activation is ongoing in the laboratories of Drs. Horowitz, McDonell, Pendergast, and Wang. Dr. Gar cia-Blanco has focused his research on the mechanism and regulation of pre-mRNA processing in mammalian cells and in Drosophila melanogaster. The regulation of DNA synthesis as an integral decision of cycling cells is investigated in Xenopous laevis oocytes by Dr. Kornbluth. All of these projects require precise quantification of specific macromolecules, which is best done using radiolabel and PhosphorImager technology. This dynamic and productive group of investigators needs this technology in order to continue an outstanding program of research. In the proposal it is explained how this instrument will benefit the group and the institution.
