An N-terminal motif in NLR immune receptors is functionally conserved across distantly related plant species

  1. Hiroaki Adachi
  2. Mauricio Contreras
  3. Adeline Harant
  4. Chih-hang Wu
  5. Lida Derevnina
  6. Toshiyuki Sakai
  7. Cian Duggan
  8. Eleonora Moratto
  9. Tolga O Bozkurt
  10. Abbas Maqbool
  11. Joe Win
  12. Sophien Kamoun  Is a corresponding author
  1. The Sainsbury Laboratory, University of East Anglia, United Kingdom
  2. Imperial College London, United Kingdom

Abstract

The molecular codes underpinning the functions of plant NLR immune receptors are poorly understood. We used in vitro Mu transposition to generate a random truncation library and identify the minimal functional region of NLRs. We applied this method to NRC4—a helper NLR that functions with multiple sensor NLRs within a Solanaceae receptor network. This revealed that the NRC4 N-terminal 29 amino acids are sufficient to induce hypersensitive cell death. This region is defined by the consensus MADAxVSFxVxKLxxLLxxEx (MADA motif) that is conserved at the N-termini of NRC family proteins and ~20% of coiled-coil (CC)-type plant NLRs. The MADA motif matches the N-terminal a1 helix of Arabidopsis NLR protein ZAR1, which undergoes a conformational switch during resistosome activation. Immunoassays revealed that the MADA motif is functionally conserved across NLRs from distantly related plant species. NRC-dependent sensor NLRs lack MADA sequences indicating that this motif has degenerated in sensor NLRs over evolutionary time.

Data availability

All sequence data used for bioinformatic and phylogenetic analyses are included in the manuscript and supporting files

The following previously published data sets were used

Article and author information

Author details

  1. Hiroaki Adachi

    The Sainsbury Laboratory, University of East Anglia, Norwich, United Kingdom
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-7184-744X
  2. Mauricio Contreras

    The Sainsbury Laboratory, University of East Anglia, Norwich, United Kingdom
    Competing interests
    No competing interests declared.
  3. Adeline Harant

    The Sainsbury Laboratory, University of East Anglia, Norwich, United Kingdom
    Competing interests
    No competing interests declared.
  4. Chih-hang Wu

    The Sainsbury Laboratory, University of East Anglia, Norwich, United Kingdom
    Competing interests
    Chih-hang Wu, SK, LD and CH-W filed a patent on NRCs.(WO/2019/108619).
  5. Lida Derevnina

    The Sainsbury Laboratory, University of East Anglia, Norwich, United Kingdom
    Competing interests
    Lida Derevnina, SK, LD and CH-W filed a patent on NRCs. (WO/2019/108619).
  6. Toshiyuki Sakai

    The Sainsbury Laboratory, University of East Anglia, Norwich, United Kingdom
    Competing interests
    No competing interests declared.
  7. Cian Duggan

    Department of Life Sciences, Imperial College London, London, United Kingdom
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-7302-7472
  8. Eleonora Moratto

    Department of Life Sciences, Imperial College London, London, United Kingdom
    Competing interests
    No competing interests declared.
  9. Tolga O Bozkurt

    Department of Life Sciences, Imperial College London, London, United Kingdom
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-0507-6875
  10. Abbas Maqbool

    The Sainsbury Laboratory, University of East Anglia, Norwich, United Kingdom
    Competing interests
    No competing interests declared.
  11. Joe Win

    The Sainsbury Laboratory, University of East Anglia, Norwich, United Kingdom
    Competing interests
    No competing interests declared.
  12. Sophien Kamoun

    The Sainsbury Laboratory, University of East Anglia, Norwich, United Kingdom
    For correspondence
    sophien.kamoun@tsl.ac.uk
    Competing interests
    Sophien Kamoun, SK, LD and CH-W filed a patent on NRCs. SK receives funding from industry on NLR biology.(WO/2019/108619).
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-0290-0315

Funding

Gatsby Charitable Foundation

  • Sophien Kamoun

Biotechnology and Biological Sciences Research Council

  • Sophien Kamoun

European Research Council

  • Sophien Kamoun

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Copyright

© 2019, Adachi et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

Metrics

  • 9,052
    views
  • 1,364
    downloads
  • 181
    citations

Views, downloads and citations are aggregated across all versions of this paper published by eLife.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

  1. Hiroaki Adachi
  2. Mauricio Contreras
  3. Adeline Harant
  4. Chih-hang Wu
  5. Lida Derevnina
  6. Toshiyuki Sakai
  7. Cian Duggan
  8. Eleonora Moratto
  9. Tolga O Bozkurt
  10. Abbas Maqbool
  11. Joe Win
  12. Sophien Kamoun
(2019)
An N-terminal motif in NLR immune receptors is functionally conserved across distantly related plant species
eLife 8:e49956.
https://doi.org/10.7554/eLife.49956

Share this article

https://doi.org/10.7554/eLife.49956

Further reading

    1. Plant Biology
    Xiaoyan Liang, Caifu Jiang
    Insight

    Salt stress delays seed germination in plants by increasing the hydrolysis of arginine-derived urea.

    1. Plant Biology
    Xinyu Chen, Huasong Zou ... Xiaojing Fan
    Research Article

    Xanthomonas citri subsp. citri (Xcc), the causal agent of citrus canker, elicits canker symptoms in citrus plants because of the transcriptional activator-like (TAL) effector PthA4, which activates the expression of the citrus susceptibility gene CsLOB1. This study reports the regulation of the putative carbohydrate-binding protein gene Cs9g12620 by PthA4-mediated induction of CsLOB1 during Xcc infection. We found that the transcription of Cs9g12620 was induced by infection with Xcc in a PthA4-dependent manner. Even though it specifically bound to a putative TAL effector-binding element in the Cs9g12620 promoter, PthA4 exerted a suppressive effect on the promoter activity. In contrast, CsLOB1 bound to the Cs9g12620 promoter to activate its expression. The silencing of CsLOB1 significantly reduced the level of expression of Cs9g12620, which demonstrated that Cs9g12620 was directly regulated by CsLOB1. Intriguingly, PhtA4 interacted with CsLOB1 and exerted feedback control that suppressed the induction of expression of Cs9g12620 by CsLOB1. Transient overexpression and gene silencing revealed that Cs9g12620 was required for the optimal development of canker symptoms. These results support the hypothesis that the expression of Cs9g12620 is dynamically directed by PthA4 for canker formation through the PthA4-mediated induction of CsLOB1.