In situ structure and organisation of the type IX secretion system

Gorasia, D. G.; Chreifi, G.; Seers, C. A.; Butler, C. A.; Heath, J. E.; Glew, M. D.; McBride, M. J.; Subramanian, P.; Kjaer, A.; Jensen, G. J.; Veith, P. D.; Reynolds, E. C.

doi:10.1101/2020.05.13.094771

bioRxiv · 10.1101/2020.05.13.094771

In situ structure and organisation of the type IX secretion system

Gorasia, D. G.·Chreifi, G.·Seers, C. A.·Butler, C. A.·Heath, J. E.·Glew, M. D.·McBride, M. J.·Subramanian, P.·Kjaer, A.·Jensen, G. J.·Veith, P. D.·Reynolds, E. C.

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Abstract

The Bacteroidetes type IX secretion system (T9SS) consists of at least 19 components that translocate proteins with a type A or type B C-terminal domain (CTD) signal across the outer membrane. The overall organisation and architecture of this system including how the secretion pore (Sov) interacts with the other components is unknown. We used cryo-electron tomography to obtain the first images of the T9SS including PorK/N rings inside intact Porphyromonas gingivalis cells. Using proteomics, we identified a novel complex between Sov, PorV and PorA and showed that Sov interacts with the PorK/N rings via PorW and a new component PGN_1783. A separate complex comprising the outer membrane {beta}-barrel protein PorP, PorE, and the type B CTD protein PG1035 was also identified. Similarly, the Flavobacterium johnsoniae PorP-like protein, SprF was found bound to the major gliding motility adhesin, SprB. Based on these data, we propose cell surface anchorage for type B CTD proteins to PorP-like proteins and a unique model where the PorK/N rings function as an outer membrane barrier to maintain the close proximity of the translocon to the shuttle and attachment complexes inside the rings, ensuring the harmonized secretion and cell surface attachment of the T9SS substrates.

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Gorasia, D. G., Chreifi, G., Seers, C. A., Butler, C. A., Heath, J. E., Glew, M. D., McBride, M. J., Subramanian, P., Kjaer, A., Jensen, G. J., Veith, P. D., Reynolds, E. C.. 2020-05-14. In situ structure and organisation of the type IX secretion system. https://doi.org/10.1101/2020.05.13.094771

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