Article

Mechanism of host substrate acetylation by a YopJ family effector

  • Nature Plants 3, Article number: 17115 (2017)
  • doi:10.1038/nplants.2017.115
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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.

Author information

Author notes

    • Zhi-Min Zhang
    • , Ka-Wai Ma
    •  & Linfeng Gao

    These authors contributed equally to this work.

Affiliations

  1. Department of Biochemistry, University of California, Riverside, California 92521, USA

    • Zhi-Min Zhang
    •  & Jikui Song
  2. Department of Plant Pathology and Microbiology, University of California, Riverside, California 92521, USA

    • Ka-Wai Ma
    • , Simon Schwizer
    •  & Wenbo Ma
  3. Environmental Toxicology Program, University of California, Riverside, California 92521, USA

    • Linfeng Gao
    •  & Jikui Song
  4. High Magnetic Field Laboratory, Chinese Academy of Sciences, Anhui 230031, China

    • Zhenquan Hu
  5. Center for Plant Cell Biology, University of California, Riverside, California 92521, USA

    • Wenbo Ma
  6. Institute of Integrative Genome Biology, University of California, Riverside, California 92521, USA

    • Wenbo Ma

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Contributions

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.

Competing interests

The authors declare no competing financial interest.

Corresponding authors

Correspondence to Wenbo Ma or Jikui Song.

Supplementary information

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    Supplementary Information

    Supplementary Figures 1–9, Supplementary Tables 1 and 2.