Journal Article

PUBDB-2024-00587

http://join2-wiki.gsi.de/foswiki/pub/Main/Artwork/join2_logo100x88.png Helical reconstruction of Salmonella and Shigella needle filaments attached to type 3 basal bodies

Kotov, V.Extern*CSSB*DESY* ; Lunelli, M.Extern*CSSB* ; Wald, J.CSSB*DESY* ; Kolbe, M. (Corresponding author)CSSB*EMBL* ; Marlovits, T. (Corresponding author)CSSB*DESY*

2021
Elsevier Amsterdam [u.a.]

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Please use a persistent id in citations: doi:10.1016/j.bbrep.2021.101039  doi:10.3204/PUBDB-2024-00587

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.

Note: Part of this work was performed at the EM facility at CSSB, supported by UHH and DFG grant numbers (INST 152/772–1|152/774–1|152/775–1|152/776–1|152-777-1 FUGG).

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Contributing Institute(s):

1. CSSB-HZI-MK (CSSB-HZI-MK)
2. CSSB-UKE-TM (CSSB-UKE-TM)

Research Program(s):

1. 633 - Life Sciences – Building Blocks of Life: Structure and Function (POF4-633) (POF4-633)
2. FUTURE T3SS - Bacterial effector secretion: Function and Architecture of the Type 3 Secretion System (311374) (311374)
3. iNEXT - Infrastructure for NMR, EM and X-rays for translational research (653706) (653706)

Experiment(s):

1. No specific instrument

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