Abstract
The Yersinia outer protein J (YopJ) family of bacterial effectors depends on a novel acetyltransferase domain to acetylate signalling proteins from plant and animal hosts. However, the underlying mechanism is unclear. Here, we report the crystal structures of PopP2, a YopJ effector produced by the plant pathogen Ralstonia solanacearum, in complex with inositol hexaphosphate (InsP6), acetyl-coenzyme A (AcCoA) and/or substrate Resistance to Ralstonia solanacearum 1 (RRS1-R)WRKY. PopP2 recognizes the WRKYGQK motif of RRS1-RWRKY to position a targeted lysine in the active site for acetylation. Importantly, the PopP2–RRS1-RWRKY association is allosterically regulated by InsP6 binding, suggesting a previously unidentified role of the eukaryote-specific cofactor in substrate interaction. Furthermore, we provide evidence for the reaction intermediate of PopP2-mediated acetylation, an acetyl-cysteine covalent adduct, lending direct support to the ‘ping-pong’-like catalytic mechanism proposed for YopJ effectors. Our study provides critical mechanistic insights into the virulence activity of YopJ class of acetyltransferases.
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Acknowledgements
This work was supported by NIH (1R35GM119721), Kimmel Scholar Award from Sidney Kimmel Foundation for Cancer Research and March of Dimes Foundation (1-FY15-345) to J.S., and grants from US NSF (IOS no. 0847870) and the USDA Agriculture Experimental Station Funding (CA-R-PPA-5075-H) to W.M. We would like to thank staff members at the Advanced Light Source, Lawrence Berkeley National Laboratory and Advanced Photon Source, Argonne National Laboratory for access to X-ray beamlines.
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Z-M.Z. and L.G. determined the crystal structures of PopP2 complexes and conducted ITC assays. J.S. and L.G. performed NMR analysis. K-W.M. and S.S. performed in vitro acetylation assays and in vivo functional analyses. Z.H. performed computational analysis. W.M. and J.S. designed and organized the study, Z.-M.Z., K-W.M., W.M. and J.S. prepared the manuscript.
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Zhang, ZM., Ma, KW., Gao, L. et al. Mechanism of host substrate acetylation by a YopJ family effector. Nature Plants 3, 17115 (2017). https://doi.org/10.1038/nplants.2017.115
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DOI: https://doi.org/10.1038/nplants.2017.115
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