- Almost every process in bacteria depends on the dynamic and specific localization of large molecules within the cell. This research addresses the question: how do bacteria make sure all of these molecules are organized into their correct positions? This project will probe the molecular and biophysical basis of cellular organization, including the role of positional constraints in regulating both essential and non-essential processes. The study will be carried out by undergraduate and graduate students in a highly mentored, collaborative research environment that emphasizes learning-through-teaching experiences and promotes critical thinking by allowing students to make decisions and learn from mistakes. The research will also produce a broadly useful and powerful community research tool that will facilitate the research efforts of over a hundred other laboratories. The project will provide multi-disciplinary training for graduate and undergraduate students.The premise of this research is that the bacterial nucleoid encodes a largely overlooked reservoir of topological information, and the aim is to elucidate the mechanisms by which the nucleoid functions as a primary positional determinant in the 3D landscape of a bacterial cell. Biochemical, structural, genetic, and cell biological approaches will be employed to elucidate the role of the nucleoid in the regulation cell division. In addition, a systematized approach to gene function discovery will be undertaken to identify and characterize novel factors that interact with the nucleoid and the cell envelope to drive subcellular organization. The gene discovery pipeline developed for this study will be broadly applicable to other organisms and the ordered gene expression library will be made publicly available to accelerate gene function discovery and characterization efforts in other laboratories.