期刊
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
卷 114, 期 23, 页码 6098-6103出版社
NATL ACAD SCIENCES
DOI: 10.1073/pnas.1705823114
关键词
bacterial nanomachines; protein secretion; superresolution microscopy; bacterial pathogenesis; Salmonella virulence
资金
- National Institute of Allergy and Infectious Diseases [AI030492]
- National Institute of Diabetes and Digestive and Kidney Diseases [P30 DK45735]
- Wellcome Trust [095927/A/11/Z]
Type III protein secretion machines have evolved to deliver bacterially encoded effector proteins into eukaryotic cells. Although electron microscopy has provided a detailed view of these machines in isolation or fixed samples, little is known about their organization in live bacteria. Here we report the visualization and characterization of the Salmonella type III secretion machine in live bacteria by 2D and 3D single-molecule switching superresolution microscopy. This approach provided access to transient components of this machine, which previously could not be analyzed. We determined the subcellular distribution of individual machines, the stoichiometry of the different components of this machine in situ, and the spatial distribution of the substrates of this machine before secretion. Furthermore, by visualizing this machine in Salmonella mutants we obtained major insights into the machine's assembly. This study bridges a major resolution gap in the visualization of this nanomachine and may serve as a paradigm for the examination of other bacterially encoded molecular machines.
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