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

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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

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Article and author information

Author details

Hiroaki Adachi

The Sainsbury Laboratory, University of East Anglia, Norwich, United Kingdom

Competing interests
No competing interests declared.

"This ORCID iD identifies the author of this article:" 0000-0002-7184-744X
Mauricio Contreras

The Sainsbury Laboratory, University of East Anglia, Norwich, United Kingdom

Competing interests
No competing interests declared.
Adeline Harant

The Sainsbury Laboratory, University of East Anglia, Norwich, United Kingdom

Competing interests
No competing interests declared.
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).
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).
Toshiyuki Sakai

The Sainsbury Laboratory, University of East Anglia, Norwich, United Kingdom

Competing interests
No competing interests declared.
Cian Duggan

Department of Life Sciences, Imperial College London, London, United Kingdom

Competing interests
No competing interests declared.

"This ORCID iD identifies the author of this article:" 0000-0001-7302-7472
Eleonora Moratto

Department of Life Sciences, Imperial College London, London, United Kingdom

Competing interests
No competing interests declared.
Tolga O Bozkurt

Department of Life Sciences, Imperial College London, London, United Kingdom

Competing interests
No competing interests declared.

"This ORCID iD identifies the author of this article:" 0000-0003-0507-6875
Abbas Maqbool

The Sainsbury Laboratory, University of East Anglia, Norwich, United Kingdom

Competing interests
No competing interests declared.
Joe Win

The Sainsbury Laboratory, University of East Anglia, Norwich, United Kingdom

Competing interests
No competing interests declared.
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).

"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

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.