The movement of bacteria toward or away from chemicals (chemotaxis) is important for cell growth and survival and may also play a major role in the progression of many infectious diseases. Escherichia coli is the pre-eminent model system for understanding the mechanisms underlying transmembrane signaling, motility and cellular behavior. E. coli chemotaxis/motility pathway is one of the best-studied sensory signal transduction systems in biology, yet it remains to be fully understood at the molecular level. Our long-term goal is to elucidate the structural basis of chemotactic signaling and flagellar switchin in living cells. The overall objective of this proposal is to correlate structural changes in the chemoreceptor complex and the flagellar motor of cells in different signaling states. By imaging flagellated minicells (~ 0.2 um diameter) generated from a skinny mreB mutant, we will be able to use cryo-electron tomography to determine high-resolution structures of the receptor array and flagellar motor trapped in different signaling states within the same individual minicells. Innovative microfluidic techniques will enable us to record the behavior of these minicells. Structure and function can be observed under identical conditions in an unprecedented manner. Thus, we will be able to test hypotheses about how changes in the structure of receptor arrays and motor complexes are manifested as changes in behavior.

Public Health Relevance

Bacterial chemotaxis and motility are important for cell growth and survival, and also play major roles in the infectivity of many prokaryotic pathogens such as Helicobacter pylori, Vibrio cholerae, Treponema pallidum and Borrelia burgorferi. We develop innovative approaches combining cryo-electron tomography and microfluidics to obtain a holistic view of the signal transduction pathway in Escherichia coli at the structural and behavioral levels. This research will improve our understanding of the mechanisms underlying transmembrane signaling, motility and cellular behavior that are widely applicable to other prokaryotic and eukaryotic systems, providing new strategies for combating bacterial pathogens.

National Institute of Health (NIH)
National Institute of General Medical Sciences (NIGMS)
Research Project (R01)
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Macromolecular Structure and Function C Study Section (MSFC)
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Flicker, Paula F
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University of Texas Health Science Center Houston
Schools of Medicine
United States
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Morado, Dustin R; Hu, Bo; Liu, Jun (2016) Using Tomoauto: A Protocol for High-throughput Automated Cryo-electron Tomography. J Vis Exp :e53608
Farley, Madeline M; Hu, Bo; Margolin, William et al. (2016) Minicells, Back in Fashion. J Bacteriol 198:1186-95
Hu, Bo; Morado, Dustin R; Margolin, William et al. (2015) Visualization of the type III secretion sorting platform of Shigella flexneri. Proc Natl Acad Sci U S A 112:1047-52
Prüβ, Birgit M; Liu, Jun; Higgs, Penelope I et al. (2015) Lessons in Fundamental Mechanisms and Diverse Adaptations from the 2015 Bacterial Locomotion and Signal Transduction Meeting. J Bacteriol 197:3028-40