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Architecture of the major component of the type III secretion system export apparatus

Abrusci, Patrizia and Vergara-Irigaray, Marta and Johnson, Steven and Beeby, Morgan D. and Hendrixson, David R. and Roversi, Pietro and Friede, Miriam E. and Deane, Janet E. and Jensen, Grant J. and Tang, Christopher M. and Lea, Susan M. (2013) Architecture of the major component of the type III secretion system export apparatus. Nature Structural & Molecular Biology, 20 (1). pp. 99-104. ISSN 1545-9985. PMCID PMC3537844. https://resolver.caltech.edu/CaltechAUTHORS:20130131-161741196

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Abstract

Type III secretion systems (T3SSs) are bacterial membrane–embedded nanomachines designed to export specifically targeted proteins from the bacterial cytoplasm. Secretion through T3SS is governed by a subset of inner membrane proteins termed the 'export apparatus'. We show that a key member of the Shigella flexneri export apparatus, MxiA, assembles into a ring essential for secretion in vivo. The ring-forming interfaces are well-conserved in both nonflagellar and flagellar homologs, implying that the ring is an evolutionarily conserved feature in these systems. Electron cryo-tomography revealed a T3SS-associated cytoplasmic torus of size and shape corresponding to those of the MxiA ring aligned to the secretion channel located between the secretion pore and the ATPase complex. This defines the molecular architecture of the dominant component of the export apparatus and allows us to propose a model for the molecular mechanisms controlling secretion.


Item Type:Article
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URLURL TypeDescription
http://dx.doi.org/10.1038/nsmb.2452DOIArticle
http://www.nature.com/nsmb/journal/v20/n1/full/nsmb.2452.htmlPublisherArticle
http://rdcu.be/cnKzPublisherFree ReadCube access
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3537844/PubMed CentralArticle
ORCID:
AuthorORCID
Johnson, Steven0000-0001-9487-8583
Beeby, Morgan D.0000-0001-6413-9835
Jensen, Grant J.0000-0003-1556-4864
Additional Information:© 2012 Nature Publishing Group, a division of Macmillan Publishers Limited. Received 30 March; accepted 22 October; published online 9 December 2012. We thank N.C. Strynadka for access to InvA coordinates ahead of publication, P. Sansonetti (Institute Pasteur, Paris) for providing the polyclonal antibodies to Ipa proteins, I. Lasa (Universidad Pública de Navarra, Pamplona) for the pKO3blue plasmid, L. De Colibus, C. King and members of the Lea group for general assistance, the staff of the protein crystallography beamlines at the European Synchrotron Radiation Source, Grenoble (France) and DIAMOND facility, Didcot (UK) for assistance in data collection, and S.K. Mazmanian for use of the microaerobic chamber. P.A. is funded by grant 083599/Z/07/Z and J.E.D. by grant WT083599MA, both from the Wellcome Trust to S.M.L.; S.J. is funded by grant G0900888 from the UK Medical Research Council to S.M.L. and C.M.T.; P.R. is funded by the Oxford Martin School Vaccine Design Institute of which S.M.L. is codirector. M.V.-I. is funded by FP7 Marie Curie EIMID-IAPP-217768 grant. M.B. and G.J.J. were supported by the Howard Hughes Medical Institute. Author Contributions: J.E.D., S.J. and S.M.L. initiated the project, which P.A. later joined; S.M.L. and C.M.T. supervised the project. M.E.F. and J.E.D. designed the MxiAC expression vector and did protein expression and stability trials. P.A. performed the large-scale purification, methylation, crosslinking and SPR of MxiAC and its mutants. S.J. performed the MALS experiments. P.A. designed the ‘export apparatus’ co-expression vectors and purified the recombinant complex. P.A. crystallized MxiAC and optimized crystals for data collection, and S.M.L. soaked and handled crystals for data collection. P.A., P.R. and S.M.L. contributed to the data collection, structure determination and model building. C.M.T. and M.V.-I. designed and performed the complementation and invasion assays in S. flexneri. D.R.H. created the C. jejuni strains, and M.D.B. and G.J.J. designed and performed cryo-EM tomography. P.A., S.J. and S.M.L. analyzed data and wrote the manuscript. All authors read and approved the manuscript.
Funders:
Funding AgencyGrant Number
Wellcome Trust083599/Z/07/Z
Wellcome TrustWT083599MA
Medical Research Council (UK)G0900888
Oxford Martin School Vaccine Design InstituteUNSPECIFIED
Marie Curie FellowshipEIMID-IAPP-217768
Howard Hughes Medical Institute (HHMI)UNSPECIFIED
Issue or Number:1
PubMed Central ID:PMC3537844
Record Number:CaltechAUTHORS:20130131-161741196
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20130131-161741196
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:36726
Collection:CaltechAUTHORS
Deposited By: Jason Perez
Deposited On:01 Feb 2013 16:08
Last Modified:09 Mar 2020 13:18

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