Intellectual merit of the proposed activity Long-range regulation of transcription occurs at several levels of organization, including entire chromosomes, blocks of genes, boundaries, and enhancers. In the even-skipped (eve) gene, recently identified elements, separate from enhancers, provide: 1. appropriate overall levels of enhancer activity, 2. proper maintenance of enhancer activity, 3. boundary activity, functionally isolating eve from nearby genes (enhancer blocking). This proposal uses genetics, reverse genetics, and biochemical approaches in Drosophila to investigate the phenomenology of these elements, and to understand their mechanisms. The larger scientific goal of these studies is to understand how protein complexes interacting with regulatory DNA mediate locus-wide regulation of gene expression, including the mechanisms of transcriptional memory. This goal represents a major remaining frontier in the quest to learn how the genome directs the development and behavior of multicellular organisms, and their interactions with their environment. eve is involved in genetic networks that direct cell behaviors along specific developmental pathways. Its regulation has been well-studied, but its genetic mechanisms are less well understood. This project will identify downstream target genes, and study how Eve regulates its target genes to generate the normal output of the pathways in which it is involved. Broader impacts of the proposed activity I. Students are strongly encouraged to participate. Both graduate and undergraduate students participate in the project. Further, undergraduates who work as technical assistants are expected to actively participate in all aspects of the scientific process, and as a result, they often share authorship on publications. Post-doctoral associates receive training that prepares them to do independent research in the future. In all of these categories, women are highly represented. Because the work routinely involves collaborative efforts with other laboratories here and abroad, it promotes the research network in biology. II. In addition to involving graduate students and undergraduates directly, the work provides exciting and current examples that are incorporated into lectures for graduate students, as well as presentations in a community Outreach Program. All of these things advance discovery and understanding while promoting teaching, training, and learning. III. The results of the work are disseminated broadly through presentations at research conferences as well as in scientific publications. Aspects of the work have been included in a number of college textbooks in recent years, including Developmental Biology by Scott F. Gilbert, and Molecular Biology of the Cell, by Bruce Alberts, et al. IV. This award also facilitates a recently established Outreach Program: A. K-12 student participation generates increased excitement in biological sciences, and encourages consideration of scientific career options. B. Teacher training is included. Under-represented groups with respect to economic status and race are largely those served in the Philadelphia, PA and Camden, NJ school districts, which are the target institutions of this program.
