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Title: Three-dimensional reconstruction of the Shigella T3SS transmembrane regions reveals 12-fold symmetry and novel features throughout

Abstract

Type III secretion systems (T3SSs) mediate bacterial protein translocation into eukaryotic cells, a process essential for virulence of many Gram-negative pathogens. They are composed of a cytoplasmic secretion machinery and a base that bridges both bacterial membranes, into which a hollow, external needle is embedded. When isolated, the latter two parts are termed the 'needle complex'. An incomplete understanding of the structure of the needle complex has hampered studies of T3SS function. To estimate the stoichiometry of its components, we measured the mass of its subdomains by scanning transmission electron microscopy (STEM). We determined subunit symmetries by analysis of top and side views within negatively stained samples in low-dose transmission electron microscopy (TEM). Application of 12-fold symmetry allowed generation of a 21-25-{angstrom} resolution, three-dimensional reconstruction of the needle complex base, revealing many new features and permitting tentative docking of the crystal structure of EscJ, an inner membrane component.

Authors:
; ; ; ; ; ; ; ;
Publication Date:
Research Org.:
Brookhaven National Lab. (BNL), Upton, NY (United States)
Sponsoring Org.:
USDOE SC OFFICE OF SCIENCE (SC)
OSTI Identifier:
1040073
Report Number(s):
BNL-90079-2009-JA
Journal ID: ISSN 1545-9993; R&D Project: BO-108; KP1501010; TRN: US201210%%260
DOE Contract Number:  
DE-AC02-98CH10886
Resource Type:
Journal Article
Journal Name:
Nature Structural and Molecular Biology
Additional Journal Information:
Journal Volume: 16; Journal Issue: 5; Journal ID: ISSN 1545-9993
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; 60 APPLIED LIFE SCIENCES; CRYSTAL STRUCTURE; MACHINERY; MEMBRANES; PATHOGENS; PROTEINS; RESOLUTION; SECRETION; SHIGELLA; STOICHIOMETRY; SYMMETRY; TRANSLOCATION; TRANSMISSION ELECTRON MICROSCOPY; VIRULENCE

Citation Formats

Hodgkinson, J L, Wall, J, Horsley, A, Stabat, S, Simon, M, Johnson, S, da Fonseca, P. C. A., Morris, E P, Lea, S M, and Blocker, A J. Three-dimensional reconstruction of the Shigella T3SS transmembrane regions reveals 12-fold symmetry and novel features throughout. United States: N. p., 2009. Web. doi:10.1038/nsmb.1599.
Hodgkinson, J L, Wall, J, Horsley, A, Stabat, S, Simon, M, Johnson, S, da Fonseca, P. C. A., Morris, E P, Lea, S M, & Blocker, A J. Three-dimensional reconstruction of the Shigella T3SS transmembrane regions reveals 12-fold symmetry and novel features throughout. United States. https://doi.org/10.1038/nsmb.1599
Hodgkinson, J L, Wall, J, Horsley, A, Stabat, S, Simon, M, Johnson, S, da Fonseca, P. C. A., Morris, E P, Lea, S M, and Blocker, A J. 2009. "Three-dimensional reconstruction of the Shigella T3SS transmembrane regions reveals 12-fold symmetry and novel features throughout". United States. https://doi.org/10.1038/nsmb.1599.
@article{osti_1040073,
title = {Three-dimensional reconstruction of the Shigella T3SS transmembrane regions reveals 12-fold symmetry and novel features throughout},
author = {Hodgkinson, J L and Wall, J and Horsley, A and Stabat, S and Simon, M and Johnson, S and da Fonseca, P. C. A. and Morris, E P and Lea, S M and Blocker, A J},
abstractNote = {Type III secretion systems (T3SSs) mediate bacterial protein translocation into eukaryotic cells, a process essential for virulence of many Gram-negative pathogens. They are composed of a cytoplasmic secretion machinery and a base that bridges both bacterial membranes, into which a hollow, external needle is embedded. When isolated, the latter two parts are termed the 'needle complex'. An incomplete understanding of the structure of the needle complex has hampered studies of T3SS function. To estimate the stoichiometry of its components, we measured the mass of its subdomains by scanning transmission electron microscopy (STEM). We determined subunit symmetries by analysis of top and side views within negatively stained samples in low-dose transmission electron microscopy (TEM). Application of 12-fold symmetry allowed generation of a 21-25-{angstrom} resolution, three-dimensional reconstruction of the needle complex base, revealing many new features and permitting tentative docking of the crystal structure of EscJ, an inner membrane component.},
doi = {10.1038/nsmb.1599},
url = {https://www.osti.gov/biblio/1040073}, journal = {Nature Structural and Molecular Biology},
issn = {1545-9993},
number = 5,
volume = 16,
place = {United States},
year = {Mon May 18 00:00:00 EDT 2009},
month = {Mon May 18 00:00:00 EDT 2009}
}