Bacteria constitute the majority of the world's biomass and are responsible for most bioconversion on the planet. Bacterial pathogens, on the other hand, present major threats to human health, causing numerous infectious diseases in humans. Moreover, bacteria also serve as model organisms for us to understand fundamental biological processes, especially at the molecular and cellular levels. It is thus of paramount importance to understand how molecules coordinate and interact inside bacterial cells to support life processes. In bacteria, life processes take place in a small volume of <1

Public Health Relevance

In this project, we propose to determine a quantitative, high-resolution map of cellular architecture of E. coli with single-molecule sensitivity, nanometer-scale spatial resolution and molecular specificity of each individual gene, and to profile changes of this architecture in response to environmental conditions with a set of bioimaging and systems biology tools. This system-wide view of bacterial architecture with ultimate sensitivity and resolution will not only advance fundamental microbiology and cell biology, but may also suggest new therapeutic targets for bacteria-based infectious diseases. The new high-sensitivity, high-resolution imaging techniques and proteomic analysis tools developed here will also have broad applications to other areas of biomedical research.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM096450-04
Application #
8537949
Study Section
Special Emphasis Panel (ZRG1-BCMB-A (51))
Program Officer
Lewis, Catherine D
Project Start
2010-09-30
Project End
2015-08-31
Budget Start
2013-09-01
Budget End
2014-08-31
Support Year
4
Fiscal Year
2013
Total Cost
$608,120
Indirect Cost
$240,900
Name
Harvard University
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
082359691
City
Cambridge
State
MA
Country
United States
Zip Code
02138
Wang, Siyuan; Moffitt, Jeffrey R; Dempsey, Graham T et al. (2014) Characterization and development of photoactivatable fluorescent proteins for single-molecule-based superresolution imaging. Proc Natl Acad Sci U S A 111:8452-7
Zhao, Ziqing W; Roy, Rahul; Gebhardt, J Christof M et al. (2014) Spatial organization of RNA polymerase II inside a mammalian cell nucleus revealed by reflected light-sheet superresolution microscopy. Proc Natl Acad Sci U S A 111:681-6
Jia, Shu; Vaughan, Joshua C; Zhuang, Xiaowei (2014) Isotropic 3D Super-resolution Imaging with a Self-bending Point Spread Function. Nat Photonics 8:302-306
Chung, Jean-Ju; Shim, Sang-Hee; Everley, Robert A et al. (2014) Structurally distinct Ca(2+) signaling domains of sperm flagella orchestrate tyrosine phosphorylation and motility. Cell 157:808-22
Chong, Shasha; Chen, Chongyi; Ge, Hao et al. (2014) Mechanism of transcriptional bursting in bacteria. Cell 158:314-26
Deindl, Sebastian; Hwang, William L; Hota, Swetansu K et al. (2013) ISWI remodelers slide nucleosomes with coordinated multi-base-pair entry steps and single-base-pair exit steps. Cell 152:442-52
Doksani, Ylli; Wu, John Y; de Lange, Titia et al. (2013) Super-resolution fluorescence imaging of telomeres reveals TRF2-dependent T-loop formation. Cell 155:345-56
Garner, Ethan C; Bernard, Remi; Wang, Wenqin et al. (2011) Coupled, circumferential motions of the cell wall synthesis machinery and MreB filaments in B. subtilis. Science 333:222-5
Wang, Wenqin; Li, Gene-Wei; Chen, Chongyi et al. (2011) Chromosome organization by a nucleoid-associated protein in live bacteria. Science 333:1445-9