AvrSr27 is a zinc-bound effector with a modular structure important for immune recognition

Outram, M. A.; Chen, J.; Broderick, S.; Li, Z.; Aditya, S.; Tasneem, N.; Arndell, T.; Blundell, C.; Ericsson, D. J.; Figueroa, M.; Sperschneider, J.; Dodds, P.; Williams, S. J.

doi:10.1101/2023.11.21.567997

bioRxiv · 10.1101/2023.11.21.567997

AvrSr27 is a zinc-bound effector with a modular structure important for immune recognition

Outram, M. A.·Chen, J.·Broderick, S.·Li, Z.·Aditya, S.·Tasneem, N.·Arndell, T.·Blundell, C.·Ericsson, D. J.·Figueroa, M.·Sperschneider, J.·Dodds, P.·Williams, S. J.

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Abstract

Stem rust, caused by the fungal pathogen Puccinia graminis f. sp.tritici (Pgt) is a major threat for wheat production and global food security. Central to the success of Pgt is the secretion of proteinaceous effectors that promote infection and colonisation, while immunity in wheat is driven by receptor-mediated recognition of these effectors resulting in pathogen avirulence. Here, we report the crystal structure of the cysteine-rich effector AvrSr27, the third experimentally derived structure of a Pgt effector. The AvrSr27 structure reveals a novel {beta}-strand rich modular fold consisting of two structurally similar domains and confirms the poor prediction we obtained from the AlphaFold2-derived model. The highly prevalent cysteine residues within the protein facilitate the co-ordination of 4 zinc molecules. Utilising the structure, we show that the N-terminal domain of AvrSr27 is sufficient for immune recognition and interaction by Sr27. The 7-cys motif sequence in each AvrSr27 domain, which facilitates zinc binding, was also found in two haustorially-expressed, structurally homologous Pgt proteins. Remarkably, despite relatively low sequence identity, we show that these proteins can associate with Sr27 and trigger cell death in heterologous systems and wheat protoplasts, albeit weaker than AvrSr27. Collectively, our findings have important implications for the field embarking on bespoke engineering of immunity receptors as solutions to plant disease.

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Outram, M. A., Chen, J., Broderick, S., Li, Z., Aditya, S., Tasneem, N., Arndell, T., Blundell, C., Ericsson, D. J., Figueroa, M., Sperschneider, J., Dodds, P., Williams, S. J.. 2023-11-21. AvrSr27 is a zinc-bound effector with a modular structure important for immune recognition. https://doi.org/10.1101/2023.11.21.567997

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