Characterisation of ShigellaSpa33 and ThermotogaFliM/N reveals a new model for C-ring assembly in T3SS

Melanie A. Mcdowell; Julien Marcoux; Gareth Mcvicker; Steven Johnson; Yu Hang Fong; Rebecca Stevens; Lesley A.H. Bowman; Matteo T. Degiacomi; Jun Yan; Adam Wise; Miriam E. Friede; Justin L.P. Benesch; Janet E Deane; Christoph M. Tang; Carol Vivien Robinson; Susan M. Lea

doi:10.1111/mmi.13267

Characterisation of ShigellaSpa33 and ThermotogaFliM/N reveals a new model for C-ring assembly in T3SS

Melanie A. Mcdowell, Julien Marcoux, Gareth Mcvicker, Steven Johnson, Yu Hang Fong, Rebecca Stevens, Lesley A.H. Bowman, Matteo T. Degiacomi, Jun Yan, Adam Wise, Miriam E. Friede, Justin L.P. Benesch, Janet E Deane, Christoph M. Tang, Carol Vivien Robinson, Susan M. Lea

Research output: Contribution to journal › Article › Research › peer-review

64 Citations (Scopus)

Abstract

Flagellar type III secretion systems (T3SS) contain an essential cytoplasmic-ring (C-ring) largely composed of two proteins FliM and FliN, whereas an analogous substructure for the closely related non-flagellar (NF) T3SS has not been observed in situ. We show that the spa33 gene encoding the putative NF-T3SS C-ring component in Shigella flexneri is alternatively translated to produce both full-length (Spa33-FL) and a short variant (Spa33-C), with both required for secretion. They associate in a 1:2 complex (Spa33-FL/C₂) that further oligomerises into elongated arrays in vitro. The structure of Spa33-C₂ and identification of an unexpected intramolecular pseudodimer in Spa33-FL reveal a molecular model for their higher order assembly within NF-T3SS. Spa33-FL and Spa33-C are identified as functional counterparts of a FliM-FliN fusion and free FliN respectively. Furthermore, we show that Thermotoga maritimaFliM and FliN form a 1:3 complex structurally equivalent to Spa33-FL/C₂, allowing us to propose a unified model for C-ring assembly by NF-T3SS and flagellar-T3SS.

Original language	English
Pages (from-to)	749-766
Number of pages	18
Journal	Molecular Microbiology
Volume	99
Issue number	4
DOIs	https://doi.org/10.1111/mmi.13267
Publication status	Published - Feb 2016
Externally published	Yes

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Mcdowell, M. A., Marcoux, J., Mcvicker, G., Johnson, S., Fong, Y. H., Stevens, R., Bowman, L. A. H., Degiacomi, M. T., Yan, J., Wise, A., Friede, M. E., Benesch, J. L. P., Deane, J. E., Tang, C. M., Robinson, C. V., & Lea, S. M. (2016). Characterisation of ShigellaSpa33 and ThermotogaFliM/N reveals a new model for C-ring assembly in T3SS. Molecular Microbiology, 99(4), 749-766. https://doi.org/10.1111/mmi.13267

@article{1f65852485dc431384ad7b8ee7c2ebfd,

title = "Characterisation of ShigellaSpa33 and ThermotogaFliM/N reveals a new model for C-ring assembly in T3SS",

abstract = "Flagellar type III secretion systems (T3SS) contain an essential cytoplasmic-ring (C-ring) largely composed of two proteins FliM and FliN, whereas an analogous substructure for the closely related non-flagellar (NF) T3SS has not been observed in situ. We show that the spa33 gene encoding the putative NF-T3SS C-ring component in Shigella flexneri is alternatively translated to produce both full-length (Spa33-FL) and a short variant (Spa33-C), with both required for secretion. They associate in a 1:2 complex (Spa33-FL/C2) that further oligomerises into elongated arrays in vitro. The structure of Spa33-C2 and identification of an unexpected intramolecular pseudodimer in Spa33-FL reveal a molecular model for their higher order assembly within NF-T3SS. Spa33-FL and Spa33-C are identified as functional counterparts of a FliM-FliN fusion and free FliN respectively. Furthermore, we show that Thermotoga maritimaFliM and FliN form a 1:3 complex structurally equivalent to Spa33-FL/C2, allowing us to propose a unified model for C-ring assembly by NF-T3SS and flagellar-T3SS.",

author = "Mcdowell, \{Melanie A.\} and Julien Marcoux and Gareth Mcvicker and Steven Johnson and Fong, \{Yu Hang\} and Rebecca Stevens and Bowman, \{Lesley A.H.\} and Degiacomi, \{Matteo T.\} and Jun Yan and Adam Wise and Friede, \{Miriam E.\} and Benesch, \{Justin L.P.\} and Deane, \{Janet E\} and Tang, \{Christoph M.\} and Robinson, \{Carol Vivien\} and Lea, \{Susan M.\}",

year = "2016",

month = feb,

doi = "10.1111/mmi.13267",

language = "English",

volume = "99",

pages = "749--766",

journal = "Molecular Microbiology",

issn = "0950-382X",

publisher = "Wiley-Blackwell",

number = "4",

}

Mcdowell, MA, Marcoux, J, Mcvicker, G, Johnson, S, Fong, YH, Stevens, R, Bowman, LAH, Degiacomi, MT, Yan, J, Wise, A, Friede, ME, Benesch, JLP, Deane, JE, Tang, CM, Robinson, CV & Lea, SM 2016, 'Characterisation of ShigellaSpa33 and ThermotogaFliM/N reveals a new model for C-ring assembly in T3SS', Molecular Microbiology, vol. 99, no. 4, pp. 749-766. https://doi.org/10.1111/mmi.13267

TY - JOUR

T1 - Characterisation of ShigellaSpa33 and ThermotogaFliM/N reveals a new model for C-ring assembly in T3SS

AU - Mcdowell, Melanie A.

AU - Marcoux, Julien

AU - Mcvicker, Gareth

AU - Johnson, Steven

AU - Fong, Yu Hang

AU - Stevens, Rebecca

AU - Bowman, Lesley A.H.

AU - Degiacomi, Matteo T.

AU - Yan, Jun

AU - Wise, Adam

AU - Friede, Miriam E.

AU - Benesch, Justin L.P.

AU - Deane, Janet E

AU - Tang, Christoph M.

AU - Robinson, Carol Vivien

AU - Lea, Susan M.

PY - 2016/2

Y1 - 2016/2

N2 - Flagellar type III secretion systems (T3SS) contain an essential cytoplasmic-ring (C-ring) largely composed of two proteins FliM and FliN, whereas an analogous substructure for the closely related non-flagellar (NF) T3SS has not been observed in situ. We show that the spa33 gene encoding the putative NF-T3SS C-ring component in Shigella flexneri is alternatively translated to produce both full-length (Spa33-FL) and a short variant (Spa33-C), with both required for secretion. They associate in a 1:2 complex (Spa33-FL/C2) that further oligomerises into elongated arrays in vitro. The structure of Spa33-C2 and identification of an unexpected intramolecular pseudodimer in Spa33-FL reveal a molecular model for their higher order assembly within NF-T3SS. Spa33-FL and Spa33-C are identified as functional counterparts of a FliM-FliN fusion and free FliN respectively. Furthermore, we show that Thermotoga maritimaFliM and FliN form a 1:3 complex structurally equivalent to Spa33-FL/C2, allowing us to propose a unified model for C-ring assembly by NF-T3SS and flagellar-T3SS.

AB - Flagellar type III secretion systems (T3SS) contain an essential cytoplasmic-ring (C-ring) largely composed of two proteins FliM and FliN, whereas an analogous substructure for the closely related non-flagellar (NF) T3SS has not been observed in situ. We show that the spa33 gene encoding the putative NF-T3SS C-ring component in Shigella flexneri is alternatively translated to produce both full-length (Spa33-FL) and a short variant (Spa33-C), with both required for secretion. They associate in a 1:2 complex (Spa33-FL/C2) that further oligomerises into elongated arrays in vitro. The structure of Spa33-C2 and identification of an unexpected intramolecular pseudodimer in Spa33-FL reveal a molecular model for their higher order assembly within NF-T3SS. Spa33-FL and Spa33-C are identified as functional counterparts of a FliM-FliN fusion and free FliN respectively. Furthermore, we show that Thermotoga maritimaFliM and FliN form a 1:3 complex structurally equivalent to Spa33-FL/C2, allowing us to propose a unified model for C-ring assembly by NF-T3SS and flagellar-T3SS.

UR - https://www.scopus.com/pages/publications/84958049794

U2 - 10.1111/mmi.13267

DO - 10.1111/mmi.13267

M3 - Article

C2 - 26538516

AN - SCOPUS:84958049794

SN - 0950-382X

VL - 99

SP - 749

EP - 766

JO - Molecular Microbiology

JF - Molecular Microbiology

IS - 4

ER -

Characterisation of ShigellaSpa33 and ThermotogaFliM/N reveals a new model for C-ring assembly in T3SS

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