In plants and animals, nucleotide-binding leucine-rich repeat (NLR) proteins serve as intracellular immune receptors. Defence signalling by NLRs often requires the formation of NLR heteropairs. Our knowledge of the molecular mechanism regulating this process is limited. In a reverse genetic screen to identify the partner of the Arabidopsis typical NLR, SUPRESSOR OF NPR1, CONSTITUTIVE 1 (SNC1), we discovered three NLRs that are redundantly required for SNC1-mediated defence, which were named SIDEKICK SNC1 1 (SIKIC1), SIKIC2 and SIKIC3. Immunoprecipitation–mass spectrometry analyses revealed that SIKIC2 physically associates with SNC1. We also uncovered that the protein level of SIKIC2 is under the control of two previously uncharacterized redundant E3 ubiquitin ligases MUSE1 and MUSE2. As SNC1 accumulation has previously been shown to be regulated by the E3 ubiquitin ligase SCFCPR1, this report provides evidence that the homeostasis of individual components of partnered typical NLRs is subjected to differential regulation via ubiquitin-mediated protein degradation.
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We thank Q. Xie (Institute of Genetics and Developmental Biology, Chinese Academy of Sciences) for sharing the pET28-AtUBA2-His construct, X. Deng (Peking University/Yale University) for sharing the pGEX-GST-AtUBC8 construct for protein expression, J. Stuttmann (Martin Luther University, Halle-Wittenberg) for sharing the seeds of N. benthaniana eds1 and K. Kirchsteiger and M. Estelle (UCSD) for offering insights in the in vitro ubiquitination assay. Y. Li (Institute of Genetics and Developmental Biology, Chinese Academy of Sciences) is thanked for Illumina sequencing data analyses. Y. Qiu (University of British Columbia) is thanked for helping with the phylogenetic analyses of MUSE1. We are grateful to N. Zheng and W. Xing (University of Washington) for enlightening discussions about MUSE1 secondary structure prediction and E3 functions. The Arabidopsis Biological Resource Center is acknowledged for providing the muse1-7 and muse2-1 knockout T-DNA mutant seeds. This work was financially supported by the Natural Sciences and Engineering Research Council (NSERC) Discovery program of Canada and Canadian Foundation for Innovation (CFI) (to X.L.), the Chinese Scholarship Council (to Y.W., Y.H., S.X. and X.C.), the Dewar Memorial fund (to X.L.), NSERC Graduate scholarships (to K.A. and K.C.M.J.) and the UBC Four-Year Fellowship (to F.X., K.C.M.J. and O.X.D.).
The authors declare no competing interests.
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Dong, O.X., Ao, K., Xu, F. et al. Individual components of paired typical NLR immune receptors are regulated by distinct E3 ligases. Nature Plants 4, 699–710 (2018). https://doi.org/10.1038/s41477-018-0216-8
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