English
 
User Manual Privacy Policy Disclaimer Contact us
  Advanced SearchBrowse

Item

ITEM ACTIONSEXPORT
  The common structural architecture of Shigella flexneri and Salmonella typhimurium Type Three Secretion needles.

Demers, J. P., Sgourakis, N. G., Gupta, R., Loquet, A., Giller, K., Riedel, D., et al. (2013). The common structural architecture of Shigella flexneri and Salmonella typhimurium Type Three Secretion needles. PLoS Pathogens, 9(3): e1003245. doi:10.1371/journal.ppat.1003245.

Item is

Basic

show hide
Item Permalink: http://hdl.handle.net/11858/00-001M-0000-000E-FC74-7 Version Permalink: http://hdl.handle.net/11858/00-001M-0000-0027-C83C-4
Genre: Journal Article

Files

show Files
hide Files
:
1744109.pdf (Publisher version), 6MB
Name:
1744109.pdf
Description:
-
Visibility:
Public
MIME-Type / Checksum:
application/pdf / [MD5]
Technical Metadata:
Copyright Date:
-
Copyright Info:
-
License:
-

Creators

show
hide
 Creators:
Demers, J. P.1, Author              
Sgourakis, N. G., Author
Gupta, R., Author
Loquet, A.1, Author              
Giller, K.2, Author              
Riedel, D.3, Author              
Laube, B., Author
Kolbe, M., Author
Baker, D., Author
Becker, S.2, Author              
Lange, A.1, Author              
Affiliations:
1Research Group of Solid-State NMR, MPI for biophysical chemistry, Max Planck Society, ou_persistent35              
2Department of NMR-Based Structural Biology, MPI for biophysical chemistry, Max Planck Society, ou_578567              
3Facility for Electron Microscopy, MPI for biophysical chemistry, Max Planck Society, ou_578615              

Content

show
hide
Free keywords: -
 Abstract: The Type Three Secretion System (T3SS), or injectisome, is a macromolecular infection machinery present in many pathogenic Gram-negative bacteria. It consists of a basal body, anchored in both bacterial membranes, and a hollow needle through which effector proteins are delivered into the target host cell. Two different architectures of the T3SS needle have been previously proposed. First, an atomic model of the Salmonella typhimurium needle was generated from solid-state NMR data. The needle subunit protein, PrgI, comprises a rigid-extended N-terminal segment and a helix-loop-helix motif with the N-terminus located on the outside face of the needle. Second, a model of the Shigella flexneri needle was generated from a high-resolution 7.7-angstrom cryo-electron microscopy density map. The subunit protein, MxiH, contains an N-terminal alpha-helix, a loop, another alpha-helix, a 14-residue-long beta-hairpin (Q51-Q64) and a C-terminal alpha-helix, with the N-terminus facing inward to the lumen of the needle. In the current study, we carried out solid-state NMR measurements of wild-type Shigella flexneri needles polymerized in vitro and identified the following secondary structure elements for MxiH: a rigid-extended N-terminal segment (S2-T11), an alpha-helix (L12-A38), a loop (E39-P44) and a C-terminal alpha-helix (Q45-R83). Using immunogold labeling in vitro and in vivo on functional needles, we located the N-terminus of MxiH subunits on the exterior of the assembly, consistent with evolutionary sequence conservation patterns and mutagenesis data. We generated a homology model of Shigella flexneri needles compatible with both experimental data: the MxiH solid-state NMR chemical shifts and the state-of-the-art cryoEM density map. These results corroborate the solid-state NMR structure previously solved for Salmonella typhimurium PrgI needles and establish that Shigella flexneri and Salmonella typhimurium subunit proteins adopt a conserved structure and orientation in their assembled state. Our study reveals a common structural architecture of T3SS needles, essential to understand T3SS-mediated infection and develop treatments.

Details

show
hide
Language(s): eng - English
 Dates: 2013-03-31
 Publication Status: Published online
 Pages: 11
 Publishing info: -
 Table of Contents: -
 Rev. Method: Peer
 Identifiers: DOI: 10.1371/journal.ppat.1003245
 Degree: -

Event

show

Legal Case

show

Project information

show

Source 1

show
hide
Title: PLoS Pathogens
Source Genre: Journal
 Creator(s):
Affiliations:
Publ. Info: -
Pages: - Volume / Issue: 9 (3) Sequence Number: e1003245 Start / End Page: - Identifier: -