Regulation of an organism's gene expression is essential for both normal development and response to environmental change. Transcription, the process of gene expression, is catalyzed by RNA polymerase (RNAP), which must bind to specific promoter DNA sequences to initiate transcription. Some genes are regulated by requiring binding of an activator protein before RNAP can bind. The scientific goal of this research is to define from a molecular, mechanistic, and structural perspective how the Mor protein, a member of a novel regulatory protein family, activates transcription of genes expressed during the middle phase of development of phage Mu. The experimental approach will include (i) the use of X-ray crystallography to determine the three dimensional structure of Mor protein in complexes with DNA and individual subunits of RNAP, (ii) biochemical identification of interacting regions of Mor, RNAP, and promoter DNA by cleavage with tethered Fe-BABE reagents, and (iii) determination of the effects of mutations in Mor, RNAP and promoter DNA on Mor binding, RNAP recruitment, formation of a specific promoter DNA distortion, and transcription activation. A second goal is to provide the graduate students conducting these experiments, and undergraduate and K-12 students who participate in them or hear them described, with (i) an understanding of multiple biological research technologies, (ii) training in experimental design and interpretation (iii) experience in obtaining and applying information from the scientific literature, and (iv) the excitement of making new scientific discoveries.
