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bioRxiv · 10.1101/2021.05.27.445799

The type 3 secretion system requires actin polymerization to open translocon pores

Russo, B. C.·Duncan-Lowey, J. K.·Chen, P.·Goldberg, M. B.
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Abstract

Many bacterial pathogens require a type 3 secretion system (T3SS) to establish a niche. Host contact activates bacterial T3SS assembly of a translocon pore in the host plasma membrane. Following pore formation, the T3SS docks onto the translocon pore. Docking establishes a continuous passage that enables the translocation of virulence proteins, effectors, into the host cytosol. Here we investigate the contribution of actin polymerization to T3SS-mediated translocation. Using the T3SS model organism Shigella flexneri, we show that actin polymerization is required for assembling the translocon pore in an open conformation, thereby enabling effector translocation. Opening of the pore channel is associated with a conformational change to the pore, which is dependent upon actin polymerization and a coiled-coil domain in the pore protein IpaC. An IpaC mutant is identified that shows actin polymerization-dependent pore opening is distinct from the previously described actin polymerization-dependent ruffles that are required for bacterial internalization. Moreover, actin polymerization is not required for other pore functions, including docking or pore protein insertion into the plasma membrane. Thus, activation of the T3SS is a multilayered process in which host signals are sensed by the translocon pore leading to the activation of effector translocation. Significance StatementThe type 3 secretion system (T3SS) is required for the virulence of a variety of bacteria that infect humans. The T3SS forms in the host cell membrane a pore that is the conduit for delivering virulence proteins into the cell. Here, we demonstrate that actin polymerization is necessary to convert pores into an open conformation that is competent for virulence protein delivery. We find that activation of type 3 secretion proceeds in a multistep process whereby bacteria dock onto the translocon pore and then activate secretion and delivery of virulence proteins.

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Russo, B. C., Duncan-Lowey, J. K., Chen, P., Goldberg, M. B.. 2021-05-27. The type 3 secretion system requires actin polymerization to open translocon pores. https://doi.org/10.1101/2021.05.27.445799

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