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000602312 1001_ $$0P:(DE-H253)PIP1082976$$aKotov, Vadim$$b0
000602312 245__ $$aHelical reconstruction of Salmonella and Shigella needle filaments attached to type 3 basal bodies
000602312 260__ $$aAmsterdam [u.a.]$$bElsevier$$c2021
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000602312 500__ $$aPart 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).
000602312 520__ $$aGram-negative pathogens evolved a syringe-like nanomachine, termed type 3 secretion system, to deliver protein effectors into the cytoplasm of host cells. An essential component of this system is a long helical needle filament that protrudes from the bacterial surface and connects the cytoplasms of the bacterium and the eukaryotic cell. Previous structural research was predominantly focused on reconstituted type 3 needle filaments, which lacked the biological context. In this work we introduce a facile procedure to obtain high-resolution cryo-EM structure of needle filaments attached to the basal body of type 3 secretion systems. We validate our approach by solving the structure of Salmonella PrgI filament and demonstrate its utility by obtaining the first high-resolution cryo-EM reconstruction of Shigella MxiH filament. Our work paves the way to systematic structural characterization of attached type 3 needle filaments in the context of mutagenesis studies, protein structural evolution and drug development. 
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000602312 536__ $$0G:(EU-Grant)311374$$aFUTURE T3SS - Bacterial effector secretion: Function and Architecture of the Type 3 Secretion System (311374)$$c311374$$fERC-2012-StG_20111109$$x1
000602312 536__ $$0G:(EU-Grant)653706$$aiNEXT - Infrastructure for NMR, EM and X-rays for translational research (653706)$$c653706$$fH2020-INFRAIA-2014-2015$$x2
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000602312 7001_ $$0P:(DE-H253)PIP1021127$$aLunelli, Michele$$b1
000602312 7001_ $$0P:(DE-H253)PIP1083333$$aWald, Jiri$$b2
000602312 7001_ $$0P:(DE-H253)PIP1021134$$aKolbe, Michael$$b3$$eCorresponding author
000602312 7001_ $$0P:(DE-H253)PIP1021412$$aMarlovits, Thomas$$b4$$eCorresponding author
000602312 773__ $$0PERI:(DE-600)2831046-9$$a10.1016/j.bbrep.2021.101039$$gVol. 27, p. 101039 -$$p101039 $$tBiochemistry and biophysics reports$$v27$$x2405-5808$$y2021
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