1. Helical reconstruction of Salmonella and Shigella needle filaments attached to type 3 basal bodies
- Author
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Vadim Kotov, Michele Lunelli, Jiri Wald, Michael Kolbe, and Thomas C. Marlovits
- Subjects
Type 3 secretion system ,Cryo electron microscopy ,Needle filament ,Helical reconstruction ,Shigella ,Salmonella, host-pathogen interaction ,Biology (General) ,QH301-705.5 ,Biochemistry ,QD415-436 - Abstract
Gram-negative pathogens evolved a syringe-like nanomachine, termed type 3 secretion system, to deliver protein effectors into the cytoplasm of host cells. An essential component of this system is a long helical needle filament that protrudes from the bacterial surface and connects the cytoplasms of the bacterium and the eukaryotic cell. Previous structural research was predominantly focused on reconstituted type 3 needle filaments, which lacked the biological context. In this work we introduce a facile procedure to obtain high-resolution cryo-EM structure of needle filaments attached to the basal body of type 3 secretion systems. We validate our approach by solving the structure of Salmonella PrgI filament and demonstrate its utility by obtaining the first high-resolution cryo-EM reconstruction of Shigella MxiH filament. Our work paves the way to systematic structural characterization of attached type 3 needle filaments in the context of mutagenesis studies, protein structural evolution and drug development.
- Published
- 2021
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