RUI: Characterization of Metabolic Genes Involved in Bacillus Sporulation

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During periods of starvation some bacterial species undergo a type of cell-differentiation which results in a highly resilient and dormant form called a spore. The study of sporulation is important since the spores of several bacterial species are harmful for a wide range of reasons. The main objective of this project is to discover how starving Bacillus subtilis cells still manage to have the energy to fuel sporulation. The central hypothesis is that when sporulating B. subtilis cells deplete their stores of glucose, the enzymes encoded by the mother cell metabolic gene (mmg) cluster contribute energy and material through the digestion of the cell's own fatty acids. The enzymes specified by the mmg cluster will be isolated, and their ability to participate in the necessary steps of fatty acid digestion will be examined. To determine the effects these genes have on sporulation, the energy state of sporulating mutants which lack these genes will be measured. Sporulation genes which are redundant or function in support of the mmg cluster will also be discovered by gene array experiments with these mutants. Successful completion of this research will be significant, because this knowledge will enable new investigations of other metabolic pathways used during sporulation.Broader Impact: Published results from this project will provide missing chemical data for previously unstudied genes, and will thus help improve the utility of the public genome sequence database for B. subtilis. Such data will also improve the predictive power of the overall set of genome databases, which will in turn impact the broad array of biotechnological fields which depend on genomics. Perhaps the most significant impact of this project on science and society will come from the large and diverse group of chemistry and biochemistry students that will receive training from this project at UNC-Greensboro. As they ultimately embark on their own science careers, the student participants will benefit science and society by contributing the broad range of communication and scientific skills they will learn from this project. The interdisciplinary project will advance discovery while promoting teaching and learning by inspiring students to pursue science careers in industry or graduate level research in NSF-sponsored fields. Former students from this lab are currently pursuing Ph.D's in organic chemistry and biological engineering, are working at biotechnology companies, or teaching high school science. New students will continue to perform all of the lab work, using diverse techniques including bacterial culture, DNA and protein methods, analytical chemistry, and synthetic organic chemistry. The students will also receive strong career mentoring related to professional life, graduate school, and scientific integrity. The Principal Investigator, a first-generation college graduate, will continue to recruit and mentor a diverse group of undergraduate and Masters students, which will include underrepresented groups and first-generation university students. For example, this large research group has been comprised so far of 62% female and 46% underrepresented minorities. The students will gain skills in written and oral communication through wide dissemination of their research in the scientific literature and presentations at scientific meetings. The project will also enhance the educational and research infrastructure at UNCG, not only from the student research opportunities, but also through the biochemistry lecture and laboratory courses. These courses will use this research to teach metabolic pathways along with the methods used to study them. Finally, this research will benefit society by adding needed experimental data to the theoretical gene-function assignments available in the publicly-funded B. subtilis genome database.