RNA Localization and Movement in Bacteria

Trustees Of Boston University, Boston

Investigators

Abstract

Intellectual merit: The internal organization of bacterial cells is complex. It appears that many proteins and specific regions of DNA occupy particular sub-cellular locations, and these positions can change during the cell cycle and in response to environment changes. However, much less is known about the localization and movements of RNA molecules in bacteria. The project will employ a novel, low-background fluorescent imaging method to study the biological relevance of the localization and movement of selected endogenous RNA molecules, including mRNAs and non-coding RNAs expressed from the bacterial chromosome. The following questions will be addressed: (1) How do the patterns of RNA localization in bacterial cells depend on the type of RNA molecule or the function of the encoded protein, in the case of mRNAs? (2) How are RNA synthesis, processing and localization coupled in bacterial cells? (3) What are the contributions of active transport and passive diffusion to the mechanism of RNA movement in bacteria? A strong inter-disciplinary research team will address these questions using new and traditional methods for studying RNA in live cells. The PI brings a strong molecular biology background to the team, while the co-PI contributes cutting edge quantitative live-cell imaging methods. Broader impacts The data obtained will provide a new perspective to RNA biology with broad relevance to gene regulation in bacteria. Graduate and undergraduate students will be trained in an interdisciplinary manner to address important biological problems using tools from microbiology, molecular biology and biophysics. The PI will incorporate experiments based on this study into her graduate laboratory course entitled "Introduction to Biomedical Engineering." Outreach efforts will include development of instructional modules in fluorescence microscopy suitable for high school students and their testing and refinement during the "Nanocamp" program sponsored by Boston University. Nanocamp is part of the Upward Bound program at Boston University, and is designed to inspire low-income, first-generation college students from local high schools in the Boston area to pursue science studies at Boston University. The program includes "Science Saturdays" during the school year and academically intensive six-week residential experiences in the summers. The fluorscent microscopy modules will engage students by the intrinsic appeal of visualizing individual molecules in living cells and furthermore, by involving students in developing animations based on these data for use in raising the scientific literacy of the general public.